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![Western Blot: Histone H4 [Trimethyl Lys20] Antibody [NB21-2090] - Analysis of Histone H4 K20me3 in (A) HeLa histone lysate (B) NIH/3T3 histone lysate, and (C) C.elegans lysate.](http://images.novusbio.com/fullsize/Histone-H4-[Trimethyl-Lys20]-Antibody-Western-Blot-NB21-2090-img0002.jpg "Western Blot: Histone H4 [Trimethyl Lys20] Antibody [NB21-2090] - Analysis of Histone H4 K20me3 in (A) HeLa histone lysate (B) NIH/3T3 histone lysate, and (C) C.elegans lysate.")
![Dot Blot: Histone H4 [Trimethyl Lys20] Antibody [NB21-2090] - Analysis of Histone H4 K20me3 in picomoles of peptide.](http://images.novusbio.com/fullsize/Histone-H4-[Trimethyl-Lys20]-Antibody-Dot-Blot-NB21-2090-img0001.jpg)
![Simple Western: Histone H4 [Trimethyl Lys20] Antibody [NB21-2090] - Lane view shows a specific band for Histone H4 in 0.5 mg/ml of Hela Nuclear lysate. This experiment was performed under reducing A246 conditions using the 12-230 kDa separation system.](http://images.novusbio.com/fullsize/Histone-H4-[Trimethyl-Lys20]-Antibody-Simple-Western-NB21-2090-img0003.jpg)
![Western Blot: Histone H4 [Trimethyl Lys20] Antibody [NBP2-42862] - Analysis of A. 30 ug HeLa whole cell lysate/extract B. 30 ug HeLa nuclear lysate/extract 15 % SDS-PAGE Histone H4 (tri methyl Lys20) antibody dilution: 1:2000](http://images.novusbio.com/fullsize/Histone-H4-[Trimethyl-Lys20]-Antibody-Western-Blot-NBP2-42862-img0007.jpg "Western Blot: Histone H4 [Trimethyl Lys20] Antibody [NBP2-42862] - Analysis of A. 30 ug HeLa whole cell lysate/extract B. 30 ug HeLa nuclear lysate/extract 15 % SDS-PAGE Histone H4 (tri methyl Lys20) antibody dilution: 1:2000")
![Immunocytochemistry/Immunofluorescence: Histone H4 [Trimethyl Lys20] Antibody [NBP2-42862] - Analysis of HeLa cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: Histone H4 (tri methyl Lys20) protein stained by Histone H4 (tri methyl Lys20) antibody diluted at 1:500. Red: alpha Tubulin antibody [114], a cytoskeleton marker, stained by diluted at 1:500. Blue: Hoechst 33342 staining.](http://images.novusbio.com/fullsize/Histone-H4-[Trimethyl-Lys20]-Antibody-Immunocytochemistry-Immunofluorescence-NBP2-42862-img0002.jpg)
![Immunohistochemistry-Paraffin: Histone H4 [Trimethyl Lys20] Antibody [NBP2-42862] - Analysis of rat fore brain. Histone H4K20me3 (trimethyl Lys20) antibody dilution: 1:500.](http://images.novusbio.com/fullsize/Histone-H4-[Trimethyl-Lys20]-Antibody-Immunohistochemistry-Paraffin-NBP2-42862-img0006.jpg)
![Immunohistochemistry-Paraffin: Histone H4 [Trimethyl Lys20] Antibody [NBP2-42862] - Analysis of mouse duodenum. Histone H4K20me3 (trimethyl Lys20) antibody dilution: 1:500.](http://images.novusbio.com/fullsize/Histone-H4-[Trimethyl-Lys20]-Antibody-Immunohistochemistry-Paraffin-NBP2-42862-img0003.jpg)
![Immunohistochemistry-Paraffin: Histone H4 [Trimethyl Lys20] Antibody [NBP2-42862] - Analysis of mouse liver. Histone H4K20me3 (trimethyl Lys20) antibody dilution: 1:500.](http://images.novusbio.com/fullsize/Histone-H4-[Trimethyl-Lys20]-Antibody-Immunohistochemistry-Paraffin-NBP2-42862-img0004.jpg)
![Immunohistochemistry-Paraffin: Histone H4 [Trimethyl Lys20] Antibody [NBP2-42862] - Analysis of mouse muscle. Histone H4K20me3 (trimethyl Lys20) antibody dilution: 1:500.](http://images.novusbio.com/fullsize/Histone-H4-[Trimethyl-Lys20]-Antibody-Immunohistochemistry-Paraffin-NBP2-42862-img0005.jpg)
![Dot Blot: Histone H4 [Trimethyl Lys20] Antibody [NBP2-42862] - Analysis of anti-Histone H4 (tri methyl Lys20) antibody with peptide samples. Varied amount of peptide samples were spotted onto positively charged nylon membrane and blotted with Histone H4 (tri methyl Lys20) antibody 1:5000 dilution. A: Peptide samples of H4K20 un B: Peptide samples of H4K20me1 C: Peptide samples of H4K20me2 D: Peptide samples of H4K20me3.](http://images.novusbio.com/fullsize/Histone-H4-[Trimethyl-Lys20]-Antibody-Dot-Blot-NBP2-42862-img0001.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)
![Other Histone H4 [Trimethyl Lys20] Antibody Blocking Peptide](http://images.novusbio.com/fullsize/Histone-H4-[Trimethyl-Lys20]-Peptide-Chromatin-Immunoprecipitation-NB21-2090PEP-img0002.jpg "Other Histone H4 [Trimethyl Lys20] Antibody Blocking Peptide")
![Western Blot: Histone H4 [Methyl Lys20] Antibody (RM208) [NBP2-61552] - Western Blot of acid extracts of HeLa cells (1) and recombinant Histone H4 (2), using NBP2-61552, showed a band of Histone H4 trimethylated at Lysine 20 (K20me3) in HeLa cells.](http://images.novusbio.com/fullsize/Histone-H4-Methyl-Lys20-Antibody-RM208-Western-Blot-NBP2-61552-img0004.jpg "Western Blot: Histone H4 [Methyl Lys20] Antibody (RM208) [NBP2-61552] - Western Blot of acid extracts of HeLa cells (1) and recombinant Histone H4 (2), using NBP2-61552, showed a band of Histone H4 trimethylated at Lysine 20 (K20me3) in HeLa cells.")
![Multiplex Immunoassay: Histone H4 [Methyl Lys20] Antibody (RM208) [NBP2-61552] - NBP2-61552 specifically reacts to Histone H4 trimethylated at Lysine 20 (K20me3). May slightly cross react to dimethylated Lysine 20 (K20me2) when used at higher concentrations. No cross reactivity with non-modified Lysine 20, monomethylated Lysine 20 (K20me1), or other methylations in histone H4.](http://images.novusbio.com/fullsize/Histone-H4-Methyl-Lys20-Antibody-RM208-Multiplex-Immunoassay-NBP2-61552-img0005.jpg)