Note: Mouseover a species abbreviation on the product page to display the fullname.
How can we help you?
![Western Blot: Histone H3 [Trimethyl Lys27] Antibody [NBP2-59206] - Western blot was performed on whole cell (40 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the antibody against H3K27me3. The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. Observed molecular weight is ~15 kDa.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27]-Antibody-Western-Blot-NBP2-59206-img0006.jpg "Western Blot: Histone H3 [Trimethyl Lys27] Antibody [NBP2-59206] - Western blot was performed on whole cell (40 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the antibody against H3K27me3. The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. Observed molecular weight is ~15 kDa.")
![Immunocytochemistry/Immunofluorescence: Histone H3 [Trimethyl Lys27] Antibody [NBP2-59206] - HeLa cells were stained with the antibody against H3K27me3 and with DAPI. Cells were fixed with 4% formaldehyde for 10' and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H3K27me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa Fluor 488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27]-Antibody-Immunocytochemistry-Immunofluorescence-NBP2-59206-img0007.jpg)
![Chromatin Immunoprecipitation: Histone H3 [Trimethyl Lys27] Antibody [NBP2-59206] - ChIP assays were performed using human K562 cells, the antibody against H3K27me3 and optimized PCR primer pairs for qPCR. ChIP was performed with a ChIP-seq kit, using sheared chromatin from 100,000 cells. The indicated amounts of antibody were used per ChIP experiment. IgG (1 ug/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the active genes GAPDH and EIF4A2, used as negative controls, and TSH2B and MYT1, used as positive controls. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27]-Antibody-Chromatin-Immunoprecipitation-NBP2-59206-img0003.jpg)
![Western Blot: Histone H3 [Trimethyl Lys27] Antibody [NBP2-59206] - Western blot was performed on whole cell (25 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the antibody against H3K27me3 diluted 1:500 in TBS-Tween containing 5% skimmed milk. Observed molecular weight is ~15 kDa.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27]-Antibody-Western-Blot-NBP2-59206-img0001.jpg)
![Chromatin Immunoprecipitation: Histone H3 [Trimethyl Lys27] Antibody [NBP2-59206] - ChIP assays were performed using human K562 cells, the antibody against H3K27me3 and optimized PCR primer pairs for qPCR. ChIP was performed with a ChIP-seq kit, using sheared chromatin from 1 million cells. The indicated amounts of antibody were used per ChIP experiment. IgG (1 ug/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the active genes GAPDH and EIF4A2, used as negative controls, and TSH2B and MYT1, used as positive controls. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27]-Antibody-Chromatin-Immunoprecipitation-NBP2-59206-img0002.jpg)
![ELISA: Histone H3 [Trimethyl Lys27] Antibody [NBP2-59206] - To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody against H3K27me3. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be >1:1,000,000.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27]-Antibody-ELISA-NBP2-59206-img0004.jpg)
![Dot Blot: Histone H3 [Trimethyl Lys27] Antibody [NBP2-59206] - To test the cross reactivity of the antibody against H3K27me3, a Dot Blot analysis was performed with peptides containing other modifications or unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure shows a high specificity of the antibody for the modification of interest.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27]-Antibody-Dot-Blot-NBP2-59206-img0005.jpg)
![Western Blot: Histone H3 [Trimethyl Lys4] Antibody [NBP2-54610] - Western blot was performed on whole cell (25 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the antibody against H3K4me3. The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. Observed molecular weight is ~16 kDa.](http://images.novusbio.com/fullsize/Histone-3-[Tryimethyl-Lys4]-Antibody-Western-Blot-NBP2-54610-img0003.jpg "Western Blot: Histone H3 [Trimethyl Lys4] Antibody [NBP2-54610] - Western blot was performed on whole cell (25 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the antibody against H3K4me3. The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. Observed molecular weight is ~16 kDa.")
![Immunohistochemistry: Histone H3 [Trimethyl Lys4] Antibody [NBP2-54610] - Histone 3 [Tryimethyl Lys4] Antibody [NBP2-54610] - HeLa cells were stained with the antibody against H3K4me3 and with DAPI. Cells were fixed with 4% formaldehyde for 10 minutes and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H3K4me3 antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.](http://images.novusbio.com/fullsize/Histone-3-[Tryimethyl-Lys4]-Antibody-Immunohistochemistry-NBP2-54610-img0004.jpg)
![Chromatin Immunoprecipitation: Histone H3 [Trimethyl Lys4] Antibody [NBP2-54610] - Histone 3 [Tryimethyl Lys4] [Trimethyl Lys4] Antibody [NBP2-54610] - ChIP assays were performed using human HeLa cells, the antibody against H3K4me3, and optimized PCR primer pairs for qPCR. Figure shows the recovery, expressed as a percent of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that trimethylation of K4 at histone H3 is associated with the promoters of active genes.](http://images.novusbio.com/fullsize/Histone-3-Tryimethyl-Lys4-Trimethyl-Lys4-Antibody-Chromatin-Immunoprecipitation-ChIP-NBP2-54610-img0005.jpg)
![Dot Blot: Histone H3 [Trimethyl Lys4] Antibody [NBP2-54610] - Histone 3 [Tryimethyl Lys4] Antibody [NBP2-54610] - One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:10,000. Figure shows a high specificity of the antibody for the modification of interest.](http://images.novusbio.com/fullsize/Histone-3-[Tryimethyl-Lys4]-Antibody-Dot-Blot-NBP2-54610-img0002.jpg)
![ELISA: Histone H3 [Trimethyl Lys4] Antibody [NBP2-54610] - Histone 3 [Tryimethyl Lys4] [Trimethyl Lys4] Antibody [NBP2-54610] - To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody against H3K4me3. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:18,350.](http://images.novusbio.com/fullsize/Histone-3-[Tryimethyl-Lys4]-[Trimethyl-Lys4]-Antibody-ELISA-NBP2-54610-img0006.jpg)
![Protein Array: Histone H3 [Trimethyl Lys4] Antibody [NBP2-54610] - Histone 3 [Tryimethyl Lys4] [Trimethyl Lys4] Antibody [NBP2-54610] - The specificity of the antibody was further demonstrated by peptide array analyses on an array containing 384 peptides with different combinations of modifications from histone H3, H4, H2A and H2B. The antibody was used at a dilution of 1:10,000. Figure shows a high specificity for the peptides containing the H3K4me3 modification.](http://images.novusbio.com/fullsize/Histone-3-[Tryimethyl-Lys4]-[Trimethyl-Lys4]-Antibody-Protein-Array-NBP2-54610-img0007.jpg)
![Western Blot: Histone H3 [Trimethyl Lys36] Antibody [NBP2-54611] - Histone 3 [Trimethyl Lys36] Antibody [NBP2-54611] - Western blot was performed on whole cell (25 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the antibody against H3K36me3. The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. Observed molecular weight ~16 kDa.](http://images.novusbio.com/fullsize/Histone-3-[Trimethyl-Lys36]-Antibody-Western-Blot-NBP2-54611-img0001.jpg "Western Blot: Histone H3 [Trimethyl Lys36] Antibody [NBP2-54611] - Histone 3 [Trimethyl Lys36] Antibody [NBP2-54611] - Western blot was performed on whole cell (25 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the antibody against H3K36me3. The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. Observed molecular weight ~16 kDa.")
![Immunocytochemistry/Immunofluorescence: Histone H3 [Trimethyl Lys36] Antibody [NBP2-54611] - Histone 3 [Trimethyl Lys36] Antibody [NBP2-54611] - HeLa cells were stained with the antibody against H3K36me3 and with DAPI. Cells were fixed with 4% formaldehyde for 10 min. and blocked with PBS/Triton X-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H3K36me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa Fluor 488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.](http://images.novusbio.com/fullsize/Histone-3-[Trimethyl-Lys36]-Antibody-Immunofluorescence-NBP2-54611-img0003.jpg)
![Chromatin Immunoprecipitation Sequencing: Histone H3 [Trimethyl Lys36] Antibody [NBP2-54611] - ChIP assays were performed using human K562 cells, the antibody against H3K36me3 and optimized PCR primer pairs for qPCR. ChIP was performed with a ChIP-seq kit, using sheared chromatin from 100,000 cells. A titration consisting of 0.2, 0.5, 1 and 2 ug of antibody per ChIP experiment was analyzed. IgG (1 ug/IP) was used as a negative IP control. Quantitative PCR was performed with primers for the coding region of the active GAPDH and ACTB genes, used as positive controls, and for the coding region of the inactive MB gene and the Sat satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys36]-Antibody-Chromatin-Immunoprecipitation-Sequencing-NBP2-54611-img0015.jpg)
![Chromatin Immunoprecipitation: Histone H3 [Trimethyl Lys36] Antibody [NBP2-54611] - ChIP assays were performed using human HeLa cells, the antibody against H3K36me3 and optimized PCR primer pairs for qPCR. ChIP was performed with an Histone ChIP-seq kit, using sheared chromatin from 1,000,000 cells. A titration consisting of 1, 2, 5 and 10 ug of antibody per ChIP experiment was analyzed. IgG (2 ug/IP) was used as a negative IP control. Quantitative PCR was performed with primers for the coding region of the active GAPDH and ACTB genes, used as positive controls, and for the promoter and a region located 1 kb upstream of the promoter of the GAPDH gene, used as negative controls.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys36]-Antibody-Chromatin-Immunoprecipitation-NBP2-54611-img0014.jpg)
![Dot Blot: Histone H3 [Trimethyl Lys36] Antibody [NBP2-54611] - Histone 3 [Trimethyl Lys36] Antibody [NBP2-54611] - To test the cross reactivity of the antibody against H3K36me3, a Dot Blot analysis was performed with peptides containing other modifications or unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. The figure shows a high specificity of the antibody for the modification of interest.](http://images.novusbio.com/fullsize/Histone-3-[Trimethyl-Lys36]-Antibody-Dot-Blot-NBP2-54611-img0004.jpg)
![ELISA: Histone H3 [Trimethyl Lys36] Antibody [NBP2-54611] - To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody against H3K36me3. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:132,000.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys36]-Antibody-ELISA-NBP2-54611-img0012.jpg)
![Protein Array: Histone H3 [Trimethyl Lys36] Antibody [NBP2-54611] - The specificity of the antibody was demonstrated by peptide array analyses on an array containing 384 peptides with different combinations of modifications from histone H3, H4, H2A and H2B. The antibody was used at a dilution of 1:10,000. The figure shows the specificity factor, calculated as the ratio of the average intensity of all spots containing the mark, divided by the average intensity of all spots not containing the mark. The peptide array analysis shows a slight cross reaction with H4K20me3 that was not observed in dot blot.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys36]-Antibody-Protein-Array-NBP2-54611-img0013.jpg)
![Western Blot: Histone H3 [Trimethyl Lys27] Antibody [NBP2-54614] - Western blot was performed on whole cell (25 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the antibody against H3K27me3 diluted 1:500 in TBS-Tween containing 5% skimmed milk. Observed molecular weight is ~17 kDa.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27]-Antibody-Western-Blot-NBP2-54614-img0013.jpg "Western Blot: Histone H3 [Trimethyl Lys27] Antibody [NBP2-54614] - Western blot was performed on whole cell (25 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the antibody against H3K27me3 diluted 1:500 in TBS-Tween containing 5% skimmed milk. Observed molecular weight is ~17 kDa.")
![Immunocytochemistry/Immunofluorescence: Histone H3 [Trimethyl Lys27] Antibody [NBP2-54614] - Human HeLa cells were stained with the antibody against H3K27me3 and with DAPI. Cells were fixed with 4% formaldehyde for 10' and blocked with PBS/Triton X-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K27me3 antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa Fluor 488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27]-Antibody-Immunocytochemistry-Immunofluorescence-NBP2-54614-img0012.jpg)
![Chromatin Immunoprecipitation: Histone H3 [Trimethyl Lys27] Antibody [NBP2-54614] - Recovery of the nucleosomes carrying the H3K27me1, H3K27me2, H3K27me3, H3K4me3, H3K9me3 and H3K36me3 modifications and the unmodified H3K27 as determined by qPCR. The figure clearly shows the antibody is very specific in ChIP for the H3K27me3 modification.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27]-Antibody-Chromatin-Immunoprecipitation-NBP2-54614-img0015.jpg)
![Chromatin Immunoprecipitation: Histone H3 [Trimethyl Lys27] Antibody [NBP2-54614] - ChIP assays were performed using human HeLa cells, the antibody against H3K27me3 and optimized PCR primer pairs for qPCR. ChIP was performed with a ChIP-seq kit, using sheared chromatin from 1 million cells. The chromatin was spiked with a panel of in vitro assembled nucleosomes, each containing a specific lysine methylation (SNAP-ChIP K-MetStat Panel, Epicypher). A titration consisting of 0.5, 1, 2 and 5 ug of antibody per ChIP experiment was analyzed. IgG (1 ug/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the active GAPDH and EIF4A2 genes, used as negative controls, and for the inactive TSH2B and MYT1 genes, used as positive controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27]-Antibody-Chromatin-Immunoprecipitation-NBP2-54614-img0014.jpg)
![ELISA: Histone H3 [Trimethyl Lys27] Antibody [NBP2-54614] - To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K27me3. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:3,000.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27]-Antibody-ELISA-NBP2-54614-img0008.jpg)
![Dot Blot: Histone H3 [Trimethyl Lys27] Antibody [NBP2-54614] - A Dot Blot analysis was performed to test the cross reactivity of the antibody against H3K27me3 with peptides containing other modifications of histone H3 and H4 and the unmodified H3K27 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:5,000. The figure shows a high specificity of the antibody for the modification of interest. Please note that the antibody also recognizes the modification if S28 is phosphorylated.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27]-Antibody-Dot-Blot-NBP2-54614-img0009.jpg)
![Western Blot: Histone H3 [ac Lys27] Antibody [NBP2-54615] - Histone 3 [Acetylated Lys27] [ac Lys27] Antibody [NBP2-54615] - Western Blot: Histone 3 [Acetylated Lys27] [ac Lys27] Antibody [NBP2-54615] - Western blot was performed on whole cell (25 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the antibody against H3K27ac. The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. Observed molecular weight is ~16 kDa.](http://images.novusbio.com/fullsize/Histone-3-[Acetylated-Lys27]-[ac-Lys27]-Antibody-Western-Blot-NBP2-54615-img0007.jpg "Western Blot: Histone H3 [ac Lys27] Antibody [NBP2-54615] - Histone 3 [Acetylated Lys27] [ac Lys27] Antibody [NBP2-54615] - Western Blot: Histone 3 [Acetylated Lys27] [ac Lys27] Antibody [NBP2-54615] - Western blot was performed on whole cell (25 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the antibody against H3K27ac. The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. Observed molecular weight is ~16 kDa.")
![Immunocytochemistry/Immunofluorescence: Histone H3 [ac Lys27] Antibody [NBP2-54615] - HeLa cells were stained with the antibody against H3K27ac and with DAPI. Cells were fixed with 4% formaldehyde for 10 minutes and blocked with PBS/ TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H3K27ac antibody (top) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown at the bottom.](http://images.novusbio.com/fullsize/Histone-3-[Acetylated-Lys27]-[ac-Lys27]-Antibody-Immunofluorescence-NBP2-54615-img0006.jpg)
![Chromatin Immunoprecipitation: Histone H3 [ac Lys27] Antibody [NBP2-54615] - ChIP assays were performed using human K562 cells, the antibody against H3K27ac and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 100,000 cells. A titration consisting of 0.2, 0.5, 1 and 2 ug of antibody per ChIP experiment was analyzed. IgG (1 ug/IP) was used as a negative IP control. Quantitative PCR was performed with primers for the promoters of the active GAPDH and EIF4A2 genes, used as positive controls, and for the coding regions of the inactive MB and MYT1 genes, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis)](http://images.novusbio.com/fullsize/Histone-H3-ac-Lys27-Antibody-Chromatin-Immunoprecipitation-ChIP-NBP2-54615-img0010.jpg)
![Chromatin Immunoprecipitation: Histone H3 [ac Lys27] Antibody [NBP2-54615] - ChIP assays were performed using human HeLa cells, the antibody against H3K27ac and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 1,000,000 cells. A titration consisting of 1, 2, 5 and 10 ug of antibody per ChIP experiment was analyzed. IgG (2 ug/IP) was used as a negative IP control. Quantitative PCR was performed with primers for the promoters of the active EIF4A2 and ACTB genes, used as positive controls, and for the inactive TSH2B and MYT1 genes, used as negative controls.](http://images.novusbio.com/fullsize/Histone-H3-ac-Lys27-Antibody-Chromatin-Immunoprecipitation-ChIP-NBP2-54615-img0009.jpg)
![Dot Blot: Histone H3 [ac Lys27] Antibody [NBP2-54615] - Histone 3 [Acetylated Lys27] [ac Lys27] Antibody [NBP2-54615] - Dot Blot: Histone 3 [Acetylated Lys27] [ac Lys27] Antibody [NBP2-54615] - To test the cross reactivity of the antibody against H3K27ac, a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K27. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure shows a high specificity of the antibody for the modification of interest.](http://images.novusbio.com/fullsize/Histone-3-[Acetylated-Lys27]-[ac-Lys27]-Antibody-Dot-Blot-NBP2-54615-img0008.jpg)
![ELISA: Histone H3 [ac Lys27] Antibody [NBP2-54615] - To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody against H3K27ac The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:8,300.](http://images.novusbio.com/fullsize/Histone-H3-[ac-Lys27]-Antibody-ELISA-NBP2-54615-img0011.jpg)
![Western Blot: Histone H3 [Trimethyl Lys9] Antibody [NBP2-54612] - Western blot was performed on whole cell (50 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3.1, H3.2 and H4 (lane 3, 4, 5, 6 and 7, respectively) using the antibody against H3K9me3. The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. Observed molecular weight is ~17 kDa.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys9]-Antibody-Western-Blot-NBP2-54612-img0016.jpg "Western Blot: Histone H3 [Trimethyl Lys9] Antibody [NBP2-54612] - Western blot was performed on whole cell (50 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3.1, H3.2 and H4 (lane 3, 4, 5, 6 and 7, respectively) using the antibody against H3K9me3. The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. Observed molecular weight is ~17 kDa.")
![Immunocytochemistry/Immunofluorescence: Histone H3 [Trimethyl Lys9] Antibody [NBP2-54612] - HeLa cells were stained with the Diagenode antibody against H3K9me3 and with DAPI. Cells were fixed with 4% formaldehyde for 10 minutes and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H3K9me3 antibody (left) diluted 1:250 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa Fluor 488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys9]-Antibody-Immunocytochemistry-Immunofluorescence-NBP2-54612-img0017.jpg)
![Chromatin Immunoprecipitation: Histone H3 [Trimethyl Lys9] Antibody [NBP2-54612] - ChIP assays were performed using human HeLa cells, the antibody against H3K9me3 and optimized PCR primer pairs for qPCR. ChIP was performed with a ChIP-seq kit, using sheared chromatin from 1,000,000 cells. A titration consisting of 1, 2, 5 and 10 ug of antibody per ChIP experiment was analyzed. IgG (2 ug/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the active genes GAPDH and EIF4A2, used as negative controls, and for ZNF510 and the Sat2 satellite repeat, used as positive controls. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys9]-Antibody-Chromatin-Immunoprecipitation-NBP2-54612-img0012.jpg)
![Immunofluorescence: Histone H3 [Trimethyl Lys9] Antibody [NBP2-54612] - NIH 3T3 cells were stained with the antibody against H3K9me3, left or with the negative control recombinant antibody, right. The bottom panel shows counterstaining of the cells with DAPI. Image using the Biotin form of this antibody.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys9]-Antibody-Immunofluorescence-NBP2-54612-img0009.jpg)
![ELISA: Histone H3 [Trimethyl Lys9] Antibody [NBP2-54612] - To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody against H3K9me3. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:42,700.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys9]-Antibody-ELISA-NBP2-54612-img0013.jpg)
![Dot Blot: Histone H3 [Trimethyl Lys9] Antibody [NBP2-54612] - To test the cross reactivity of the antibody against H3K9me3, a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K9. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure shows a high specificity of the antibody for the modification of interest.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys9]-Antibody-Dot-Blot-NBP2-54612-img0014.jpg)
![Protein Array: Histone H3 [Trimethyl Lys9] Antibody [NBP2-54612] - The specificity of the antibody was further demonstrated by peptide array analyses on an array containing 384 peptides with different combinations of modifications from histone H3, H4, H2A and H2B. The antibody was used at a dilution of 1:5,000. Figure shows the specificity factor, calculated as the ratio of the average intensity of all spots containing the mark, divided by the average intensity of all spots not containing the mark.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys9]-Antibody-Protein-Array-NBP2-54612-img0015.jpg)
![Western Blot: Histone H3 [ac Lys14, ac Lys9] Antibody [NBP2-59181] - Histone extracts of HeLa cells (15 ug) were analysed by Western blot using the antibody directed against H3K9/14ac diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. Observed molecular weight is ~15 kDa.](http://images.novusbio.com/fullsize/Histone-H3-[ac-Lys14--ac-Lys9]-Antibody-Western-Blot-NBP2-59181-img0001.jpg "Western Blot: Histone H3 [ac Lys14, ac Lys9] Antibody [NBP2-59181] - Histone extracts of HeLa cells (15 ug) were analysed by Western blot using the antibody directed against H3K9/14ac diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. Observed molecular weight is ~15 kDa.")
![Chromatin Immunoprecipitation: Histone H3 [ac Lys14, ac Lys9] Antibody [NBP2-59181] - ChIP assays were performed using HeLa cells, the antibody against H3K9/14ac and optimized primer pairs for qPCR. ChIP was performed with a ChIP kit, using sheared chromatin from 1.5 million cells. A titration of the antibody consisting of 1, 2, 5 and 10 ug per ChIP experiment was analysed. IgG (5 ug/IP) was used as negative IP control.QPCR was performed using primers specific for the promoter of the ACTB gene as a positive control target and for exon 2 of the MB gene as a negative control target. Figure shows the recovery (the relative amount of immunoprecipitated DNA compared to input DNA). These results confirm the observation that acetylation of H3K9/14 is present at active promoters.](http://images.novusbio.com/fullsize/Histone-H3-[ac-Lys14--ac-Lys9]-Antibody-Chromatin-Immunoprecipitation-NBP2-59181-img0008.jpg)
![Chromatin Immunoprecipitation: Histone H3 [ac Lys14, ac Lys9] Antibody [NBP2-59181] - ChIP assays were performed using HeLa cells, the antibody against H3K9/14ac and optimized primer pairs for qPCR. ChIP was performed using sheared chromatin from 1.5 million cells. A titration of the antibody consisting of 1, 2, 5 and 10 ug per ChIP experiment was analysed. IgG (5 ug/IP) was used as negative IP control. QPCR was performed using primers specific for the promoter of the ACTB gene as a positive control target and for exon 2 of the MB gene as a negative control target. Figure shows the recovery (the relative amount of immunoprecipitated DNA compared to input DNA). These results confirm the observation that acetylation of H3K9/14 is present at active promoters.](http://images.novusbio.com/fullsize/Histone-H3-[ac-Lys14--ac-Lys9]-Antibody-Chromatin-Immunoprecipitation-NBP2-59181-img0004.jpg)
![Chromatin Immunoprecipitation: Histone H3 [ac Lys14, ac Lys9] Antibody [NBP2-59181] - ChIP was performed with 1 ug of the antibody against H3K9/14ac on sheared chromatin from 1 million HeLaS3 cells. IgG (2 ug/IP) was used as a negative IP control. The IP'd DNA was analysed by QPCR with optimized PCR primer pairs for the promoters of the active GAPDH and c-fos genes, used as positive control targets, and the coding region of the inactive MB gene and the Sat2 satellite repeat, used as negative control targets.](http://images.novusbio.com/fullsize/Histone-H3-[ac-Lys14--ac-Lys9]-Antibody-Chromatin-Immunoprecipitation-NBP2-59181-img0005.jpg)
![Dot Blot: Histone H3 [ac Lys14, ac Lys9] Antibody [NBP2-59181] - A Dot Blot analysis was performed to test the cross reactivity of the antibody against H3K9/14ac with peptides containing other histone modifications and the unmodified H3K9/14 sequence. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure shows a high specificity of the antibody for the modification of interest.](http://images.novusbio.com/fullsize/Histone-H3-[ac-Lys14--ac-Lys9]-Antibody-Dot-Blot-NBP2-59181-img0002.jpg)
![ELISA: Histone H3 [ac Lys14, ac Lys9] Antibody [NBP2-59181] - To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody directed against H3K9/14ac, crude serum and flow through in antigen coated wells. The antigen used was a peptie containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the purified antibody was estimated to be 1:5,900.](http://images.novusbio.com/fullsize/Histone-H3-[ac-Lys14--ac-Lys9]-Antibody-ELISA-NBP2-59181-img0003.jpg)
![Immunofluorescence: Histone H3 [ac Lys14, ac Lys9] Antibody [NBP2-59181] - Mouse NIH3T3 cells were stained with the antibody against H3K9/14ac and with DAPI. Cells were fixed with 4% formaldehyde for 10 minutes and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K9/14ac antibody (left) diluted 1:500 in blocking solution followed by an anti- rabbit antibody conjugated to Alexa Fluor 488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.](http://images.novusbio.com/fullsize/Histone-H3-[ac-Lys14-ac-Lys9]-Antibody-Immunofluorescence-NBP2-59181-img0006.jpg)
![Western Blot: Histone H3 [ac Lys9, ac Lys14] Antibody [NBP2-54617] - Western blot was performed on whole cell (25 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the antibody against H3K9/14ac. The antibody was diluted 1:500 in TBS-Tween containing 5% skimmed milk. Observed molecular weight is ~16 kDa.](http://images.novusbio.com/fullsize/Histone-H3-[ac-Lys9--ac-Lys14]-Antibody-Western-Blot-NBP2-54617-img0003.jpg "Western Blot: Histone H3 [ac Lys9, ac Lys14] Antibody [NBP2-54617] - Western blot was performed on whole cell (25 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the antibody against H3K9/14ac. The antibody was diluted 1:500 in TBS-Tween containing 5% skimmed milk. Observed molecular weight is ~16 kDa.")
![Immunocytochemistry/Immunofluorescence: Histone H3 [ac Lys9, ac Lys14] Antibody [NBP2-54617] - HeLa cells were stained with the antibody against H3K9/14ac and with DAPI. Cells were fixed with 4% formaldehyde for 10' and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H3K9/14ac antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.](http://images.novusbio.com/fullsize/Histone-H3-[ac-Lys9--ac-Lys14]-Antibody-Immunofluorescence-NBP2-54617-img0007.jpg)
![Chromatin Immunoprecipitation: Histone H3 [ac Lys9, ac Lys14] Antibody [NBP2-54617] - ChIP assays were performed using human HeLa cells, the antibody against H3K9/14ac and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 1,000,000 cells. A titration consisting of 1, 2, 5 and 10 ug of antibody per ChIP experiment was analyzed. IgG (2 ug/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the active genes GAPDH and EIF4A2, used as positive controls, and for the coding region of the inactive MB gene and the Sat2 satellite repeat, used as negative controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).](http://images.novusbio.com/fullsize/Histone-H3-ac-Lys9-ac-Lys14-Antibody-Chromatin-Immunoprecipitation-ChIP-NBP2-54617-img0001.jpg)
![ELISA: Histone H3 [ac Lys9, ac Lys14] Antibody [NBP2-54617] - To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody against H3K9/14ac. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:4,000.](http://images.novusbio.com/fullsize/Histone-H3-[ac-Lys9--ac-Lys14]-Antibody-ELISA-NBP2-54617-img0002.jpg)
![Dot Blot: Histone H3 [ac Lys9, ac Lys14] Antibody [NBP2-54617] - To test the cross reactivity of the antibody against H3K9/14ac, a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K9. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure shows a high specificity of the antibody for the modification of interest.](http://images.novusbio.com/fullsize/Histone-H3-[ac-Lys9--ac-Lys14]-Antibody-Dot-Blot-NBP2-54617-img0006.jpg)
![Protein Array: Histone H3 [ac Lys9, ac Lys14] Antibody [NBP2-54617] - The specificity of the antibody was further demonstrated by peptide array analyses on an array containing 384 peptides with different combinations of modifications from histone H3, H4, H2A and H2B. The antibody was used at a dilution of 1:2,000. Figure shows the specificity factor, calculated as the ratio of the average intensity of all spots containing the mark, divided by the average intensity of all spots not containing the mark.](http://images.novusbio.com/fullsize/Histone-H3-[ac-Lys9--ac-Lys14]-Antibody-Protein-Array-NBP2-54617-img0005.jpg)
![Western Blot: HDAC1 Antibody [NBP2-54621] - Whole cell extracts (50 ug) from HeLa cells transfected with HDAC1 siRNA (lane 2) and from an untransfected control (lane 1) were analysed by Western blot using the antibody against HDAC1 diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right (expected size: 55 kDa); the marker (in kDa) is shown on the left.](http://images.novusbio.com/fullsize/HDAC1-Antibody-Western-Blot-NBP2-54621-img0004.jpg "Western Blot: HDAC1 Antibody [NBP2-54621] - Whole cell extracts (50 ug) from HeLa cells transfected with HDAC1 siRNA (lane 2) and from an untransfected control (lane 1) were analysed by Western blot using the antibody against HDAC1 diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right (expected size: 55 kDa); the marker (in kDa) is shown on the left.")
![Immunocytochemistry/Immunofluorescence: HDAC1 Antibody [NBP2-54621] - HeLa cells were stained with the antibody against HDAC1 and with DAPI. Cells were fixed with 4% formaldehyde for 10' and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the HDAC1 antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.](http://images.novusbio.com/fullsize/HDAC1-Antibody-Immunofluorescence-NBP2-54621-img0006.jpg)
![Chromatin Immunoprecipitation: HDAC1 Antibody [NBP2-54621] - ChIP was performed with the antibody against HDAC1 on sheared chromatin from 4,000,000 HeLa cells. An antibody titration consisting of 1, 2, 5 and 10 ug per ChIP experiment was analysed. IgG (2 ug/IP) was used as negative IP control. QPCR was performed with primers specific for the EIF4A2 and GAPDH promoters, used as positive controls, and for the MYOD1 gene and Sat2 satellite repeat, used as negative controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).](http://images.novusbio.com/fullsize/HDAC1-Antibody-Chromatin-Immunoprecipitation-ChIP-NBP2-54621-img0001.jpg)
![Western Blot: HDAC1 Antibody [NBP2-54621] - Whole cell extracts (25 ug, lane 1) and nuclear extracts (25 ug, lane 2) from HeLa cells were analysed by Western blot using the antibody against HDAC1 diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right (expected size: 55 kDa); the marker (in kDa) is shown on the left.](http://images.novusbio.com/fullsize/HDAC1-Antibody-Western-Blot-NBP2-54621-img0003.jpg)
![ELISA: HDAC1 Antibody [NBP2-54621] - To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody directed against HDAC1, crude serum and flow through. The plates were coated with the peptide used for immunization of the rabbit. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:75,000.](http://images.novusbio.com/fullsize/HDAC1-Antibody-ELISA-NBP2-54621-img0002.jpg)
![Protein Array: HDAC1 Antibody [NBP2-54621] - The specificity of the antibody against HDAC1 was demonstrated using a human protein microarray containing more than 19,000 human proteins. The antibody was used at a dilution of 1:100,000. The figure shows the Z-score of the signal intensity (mean value of the duplicate spots on the array). The names of the proteins with 5 highest Z-scores are indicated at the bottom. This figure clearly shows the high specificity of the antibody for HDAC1.](http://images.novusbio.com/fullsize/HDAC1-Antibody-Protein-Array-NBP2-54621-img0005.jpg)
![Western Blot: Histone H3 [Monomethyl Lys4] Antibody [NBP2-54613] - Histone 3 [Monomethyl Lys4] Antibody [NBP2-54613] - Western blot was performed on whole cell (25 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the antibody against H3K4me1. The antibody was diluted 1:500 in TBS-Tween containing 5% skimmed milk. Observed molecular weight is ~15 kDa.](http://images.novusbio.com/fullsize/Histone-3-[Monomethyl-Lys4]-Antibody-Western-Blot-NBP2-54613-img0004.jpg "Western Blot: Histone H3 [Monomethyl Lys4] Antibody [NBP2-54613] - Histone 3 [Monomethyl Lys4] Antibody [NBP2-54613] - Western blot was performed on whole cell (25 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the antibody against H3K4me1. The antibody was diluted 1:500 in TBS-Tween containing 5% skimmed milk. Observed molecular weight is ~15 kDa.")
![Immunocytochemistry/Immunofluorescence: Histone H3 [Monomethyl Lys4] Antibody [NBP2-54613] - Histone 3 [Monomethyl Lys4] Antibody [NBP2-54613] - HeLa cells were stained with the antibody against H3K4me1 and with DAPI. Cells were fixed with 4% formaldehyde for 10' and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H3K4me1 antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa Fluor 488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.](http://images.novusbio.com/fullsize/Histone-3-[Monomethyl-Lys4]-Antibody-Immunofluorescence-NBP2-54613-img0007.jpg)
![Chromatin Immunoprecipitation: Histone H3 [Monomethyl Lys4] Antibody [NBP2-54613] - Histone 3 [Monomethyl Lys4] Antibody [NBP2-54613] - ChIP assays were performed using human K562 cells, the antibody against H3K4me1 and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 100,000 cells. A titration consisting of 1, 2, 5 and 10 ug of antibody per ChIP experiment was analyzed. IgG (1 ug/IP) was used as a negative IP control. Quantitative PCR was performed with primers for a region surrounding the ACTB and GAS2L1 gene, respectively, used as positive controls, and for the promoters of the GAPDH and EIF4A2 genes, used as negative controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).](http://images.novusbio.com/fullsize/Histone-3-Monomethyl-Lys4-Antibody-Chromatin-Immunoprecipitation-ChIP-NBP2-54613-img0008.jpg)
![Dot Blot: Histone H3 [Monomethyl Lys4] Antibody [NBP2-54613] - Histone 3 [Monomethyl Lys4] Antibody [NBP2-54613] - To test the cross reactivity of the antibody against H3K4me1, a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:5,000. Figure shows a high specificity of the antibody for the modification of interest.](http://images.novusbio.com/fullsize/Histone-3-[Monomethyl-Lys4]-Antibody-Dot-Blot-NBP2-54613-img0003.jpg)
![ELISA: Histone H3 [Monomethyl Lys4] Antibody [NBP2-54613] - Histone 3 [Monomethyl Lys4] Antibody [NBP2-54613] - To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody against H3K4me1. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:10,300.](http://images.novusbio.com/fullsize/Histone-3-[Monomethyl-Lys4]-Antibody-ELISA-NBP2-54613-img0009.jpg)
![Protein Array: Histone H3 [Monomethyl Lys4] Antibody [NBP2-54613] - Histone 3 [Monomethyl Lys4] Antibody [NBP2-54613] - The specificity of the antibody was further demonstrated by peptide array analyses on an array containing 384 peptides with different combinations of modifications from histone H3, H4, H2A and H2B. The antibody was used at a dilution of 1:2,000. Figure shows the specificity factor, calculated as the ratio of the average intensity of all spots containing the mark, divided by the average intensity of all spots not containing the mark.](http://images.novusbio.com/fullsize/Histone-3-[Monomethyl-Lys4]-Antibody-Protein-Array-NBP2-54613-img0010.jpg)
![Western Blot: Histone H4 [Trimethyl Lys20] Antibody [NBP2-59259] - Histone extracts of HeLa cells (15 ug) were analysed by Western blot using Histone H4 [Trimethyl Lys20] Antibody diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.](http://images.novusbio.com/fullsize/Histone-H4-[Trimethyl-Lys20]-Antibody-Western-Blot-NBP2-59259-img0006.jpg "Western Blot: Histone H4 [Trimethyl Lys20] Antibody [NBP2-59259] - Histone extracts of HeLa cells (15 ug) were analysed by Western blot using Histone H4 [Trimethyl Lys20] Antibody diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.")
![Immunocytochemistry/Immunofluorescence: Histone H4 [Trimethyl Lys20] Antibody [NBP2-59259] - (Top): Human osteosarcoma (U2OS) cells were stained with and DAPI. Cells were fixed with ice cold methanol for 10' and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 6A: cells were immunofluorescently labeled with (left) diluted 1:300 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. (Bottom): Staining of the cells with the after incubation of the antibody with blocking peptide, concentration: 5 ng/ul.](http://images.novusbio.com/fullsize/Histone-H4-[Trimethyl-Lys20]-Antibody-Immunocytochemistry-Immunofluorescence-NBP2-59259-img0005.jpg)
![Chromatin Immunoprecipitation: Histone H4 [Trimethyl Lys20] Antibody [NBP2-59259] - ChIP assays were performed using undifferentiated human teratocarcinoma cells (NCCIT), the antibody against H4K20me3 and optimized PCR primer sets for qPCR. Sheared chromatin from 10,000 cells was used per ChIP experiment. The antibody was tested at two different dilutions of 1:500 and 1:100. The pre-immune serum (PI) was used as a negative control. Quantitative PCR was performed using primer sets for the satellite repeat Sat2 as a positive control and for the promoter of the house keeping gene c-fos, as a negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that H4K20me3 is preferably present at heterochromatin.](http://images.novusbio.com/fullsize/Histone-H4-[Trimethyl-Lys20]-Antibody-Chromatin-Immunoprecipitation-NBP2-59259-img0004.jpg)
![Western Blot: Histone H4 [Trimethyl Lys20] Antibody [NBP2-59259] - Histone (acid) extracts of NB4 cells were analysed by Western blot using the Diagenode antibody against H4K20me3, diluted 1:750 in TBS-Tween containing 5% skimmed milk. The location of the protein of interest is indicated on the right.](http://images.novusbio.com/fullsize/Histone-H4-[Trimethyl-Lys20]-Antibody-Western-Blot-NBP2-59259-img0002.jpg)
![Dot Blot: Histone H4 [Trimethyl Lys20] Antibody [NBP2-59259] - Dot Blot was used to check the specificity of the antibody against H4K20me3 with peptides containing other modifications of histone H3 and H4. Other histone modifications include monoand dimethylation of the same lysine and mono-, di- and trimethylation of lysines 9, 27 and 36 of H3. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:1,000. Figure shows a high specificity of the antibody for the modification of interest.](http://images.novusbio.com/fullsize/Histone-H4-[Trimethyl-Lys20]-Antibody-Dot-Blot-NBP2-59259-img0001.jpg)
![ELISA: Histone H4 [Trimethyl Lys20] Antibody [NBP2-59259] - To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H4K20me3. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:700.](http://images.novusbio.com/fullsize/Histone-H4-[Trimethyl-Lys20]-Antibody-ELISA-NBP2-59259-img0003.jpg)
![Dot Blot: Histone H4 [Trimethyl Lys20] Antibody [NBP2-59259] - A Dot Blot analysis was performed to test the cross reactivity of Histone H4 [Trimethyl Lys20] Antibody with peptides containing other histone modifications and the unmodified H4K20. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure shows a high specificity of the antibody for the modification of interest](http://images.novusbio.com/fullsize/Histone-H4-[Trimethyl-Lys20]-Antibody-Dot-Blot-NBP2-59259-img0007.jpg)
![Western Blot: Histone H3 [Trimethyl Lys20] Antibody [NBP2-54620] - Western blot was performed on whole cell (25 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the antibody against H4K20me3. The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. Observed molecular weight is ~11 kDa.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys20]-Antibody-Western-Blot-NBP2-54620-img0005.jpg "Western Blot: Histone H3 [Trimethyl Lys20] Antibody [NBP2-54620] - Western blot was performed on whole cell (25 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the antibody against H4K20me3. The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. Observed molecular weight is ~11 kDa.")
![Immunocytochemistry/Immunofluorescence: Histone H3 [Trimethyl Lys20] Antibody [NBP2-54620] - HeLa cells were stained with the antibody against H4K20me3 and with DAPI. Cells were fixed with methanol and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H4K20me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa Fluor 488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys20]-Antibody-Immunofluorescence-NBP2-54620-img0006.jpg)
![Chromatin Immunoprecipitation: Histone H3 [Trimethyl Lys20] Antibody [NBP2-54620] - ChIP assays were performed using human K562 cells, the antibody against H4K20me3 and optimized PCR primer sets for qPCR. ChIP was performed on sheared chromatin from 100,000 cells. A titration of the antibody consisting of 0.2, 0.5, 1 and 2 ug per ChIP experiment was analysed. IgG (1 ug/IP) was used as negative IP control. QPCR was performed with primers for promoters of the active genes EIF4A2 and GAPDH, used as negative controls, and for ZNF510 and the Sat2 satellite repeat region used as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).](http://images.novusbio.com/fullsize/Histone-H3-Trimethyl-Lys20-Antibody-Chromatin-Immunoprecipitation-ChIP-NBP2-54620-img0001.jpg)
![ELISA: Histone H3 [Trimethyl Lys20] Antibody [NBP2-54620] - To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody directed against H4K20me3 in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:21,700.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys20]-Antibody-ELISA-NBP2-54620-img0002.jpg)
![Dot Blot: Histone H3 [Trimethyl Lys20] Antibody [NBP2-54620] - To test the cross reactivity of the antibody against H4K20me3. A Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H4K20. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure shows a high specificity of the antibody for the modification of interest.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys20]-Antibody-Dot-Blot-NBP2-54620-img0003.jpg)
![Protein Array: Histone H3 [Trimethyl Lys20] Antibody [NBP2-54620] - The specificity of the antibody was further demonstrated by peptide array analyses on an array containing 384 peptides with different combinations of modifications from histone H3, H4, H2A and H2B. The antibody was used at a dilution of 1:10,000. Figure shows the specificity factor, calculated as the ratio of the average intensity of all spots containing the mark, divided by the average intensity of all spots not containing the mark.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys20]-Antibody-Protein-Array-NBP2-54620-img0004.jpg)