| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Azide and BSA Free | NBP3-25993 | |||
| Conjugate | Catalog # | Availability | Size | Price |
|---|---|---|---|---|
| DyLight 405 | NB21-1193V | |||
| FITC | NB21-1193F | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Azide and BSA Free | NBP3-12020-0.025mg | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Azide and BSA Free | NBP3-12024-0.025mg | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Azide and BSA Free | NBP3-12020-0.2mg | |||
| Azide and BSA Free | NBP3-12020-0.025mg | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Azide and BSA Free | NBP3-12024-0.2mg | |||
| Azide and BSA Free | NBP3-12024-0.025mg | |||
![Western Blot: Histone H3 [Trimethyl Lys27] Antibody (RM175) [NBP2-61550] - Western Blot of recombinant histone H3.3 (1) and acid extracts of HeLa cells (2), using NBP2-61550, showed a band of histone H3 trimethylated at Lysine 27 (K27me3) in HeLa cells. Observed molecular weight is ~17 kDa](http://images.novusbio.com/fullsize/Histone-H3-Trimethyl-Lys27-Antibody-RM175-Western-Blot-NBP2-61550-img0005.jpg "Western Blot: Histone H3 [Trimethyl Lys27] Antibody (RM175) [NBP2-61550] - Western Blot of recombinant histone H3.3 (1) and acid extracts of HeLa cells (2), using NBP2-61550, showed a band of histone H3 trimethylated at Lysine 27 (K27me3) in HeLa cells. Observed molecular weight is ~17 kDa")
![Immunohistochemistry: Histone H3 [Trimethyl Lys27] Antibody (RM175) [NBP2-61550] - Immunohistochemistry staining of HepG2 cells using NBP2-61550.](http://images.novusbio.com/fullsize/Histone-H3-Trimethyl-Lys27-Antibody-RM175-Immunohistochemistry-NBP2-61550-img0004.jpg)
![Multiplex Immunoassay: Histone H3 [Trimethyl Lys27] Antibody (RM175) [NBP2-61550] - NBP2-61550 specifically reacts to Histone H3 trimethylated at Lysine 27 (K27me3). No cross reactivity with non-modified Lysine 27 (K27 Ctrl), monomethylated Lysine 27 (K27me1) or dimethylated Lysine 27 (K27me2), or other methylations in Histone H3.](http://images.novusbio.com/fullsize/Histone-H3-Trimethyl-Lys27-Antibody-RM175-Multiplex-Immunoassay-NBP2-61550-img0006.jpg)
![Western Blot: Histone H3 [Trimethyl Lys27] Antibody [NBP2-16840] - Various whole cell extracts (30 ug) were separated by 15% SDS-PAGE, and the membrane was blotted with Histone H3K27me3 (trimethyl Lys27) antibody diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (NBP2-19301) was used to detect the primary antibody. Observed molecular weight is ~15 kDa.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27]-Antibody-Western-Blot-NBP2-16840-img0021.jpg "Western Blot: Histone H3 [Trimethyl Lys27] Antibody [NBP2-16840] - Various whole cell extracts (30 ug) were separated by 15% SDS-PAGE, and the membrane was blotted with Histone H3K27me3 (trimethyl Lys27) antibody diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (NBP2-19301) was used to detect the primary antibody. Observed molecular weight is ~15 kDa.")
![Immunocytochemistry/Immunofluorescence: Histone H3 [Trimethyl Lys27] Antibody [NBP2-16840] - 293T cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: Histone H3K27me3 (Tri-methyl Lys27) stained by Histone H3K27me3 (Tri-methyl Lys27) antibody diluted at 1:500. Red: alpha Tubulin, a cytoskeleton marker, stained by alpha Tubulin antibody [114] diluted at 1:1000.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27]-Antibody-Immunocytochemistry-Immunofluorescence-NBP2-16840-img0027.jpg)
![Immunohistochemistry-Paraffin: Histone H3 [Trimethyl Lys27] Antibody [NBP2-16840] - Mouse intestine. Histone H3K27me3 (Tri-methyl Lys27) stained by Histone H3K27me3 (Tri-methyl Lys27) antibody diluted at 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27]-Antibody-Immunohistochemistry-Paraffin-NBP2-16840-img0026.jpg)
![Western Blot: Histone H3 [Trimethyl Lys27] Antibody [NBP2-16840] - Histone H3 [Trimethyl Lys27] Antibody U87 cells were grown under normoxic (DMEM 10% FBS, 16% O2), hypoxic (DMEM 10% FBS, 1% O2), and neurosphere conditions (DMEM/F12, B-27 supplement, growth factor (10ng/ml FGF and 20ng/ml EGF)). Cell lysate were Western blotted for H3K27me3 and Histone-H3 (loading control).](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27]-Antibody-Western-Blot-NBP2-16840-img0008.jpg)
![Immunocytochemistry/Immunofluorescence: Histone H3 [Trimethyl Lys27] Antibody [NBP2-16840] - Histone H3 [Trimethyl Lys27] Antibody Confocal immunofluorescence analysis of paraformaldehyde-fixed A431, using HIST1H3A (tri-Methyl K27) antibody (Green) at 1:500 dilution. Alpha-tubulin filaments are labeled with Alpha-tubulin antibody (Red) at 1:2000.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27]-Antibody-Immunocytochemistry-Immunofluorescence-NBP2-16840-img0005.jpg)
![Immunocytochemistry/Immunofluorescence: Histone H3 [Trimethyl Lys27] Antibody [NBP2-16840] - Immunofluorescence photomicrographs of frozen sections of mouse liver. Red: Histone H3K27me3 (trimethyl Lys27) antibody diluted at 1:200. The signal was developed using goat anti-rabbit IgG antibody Blue: Nuclear staining with Hoechst 33342.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27]-Antibody-Immunofluorescence-NBP2-16840-img0011.jpg)
![Immunohistochemistry-Paraffin: Histone H3 [Trimethyl Lys27] Antibody [NBP2-16840] - Hela xenograft, using Histon H3 (tri-Methyl K27) antibody at 1:500 dilution.Antigen Retrieval: Trilogy™ (EDTA based, pH 8.0) buffer, 15min.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27]-Antibody-Immunohistochemistry-Paraffin-NBP2-16840-img0006.jpg)
![Immunohistochemistry-Paraffin: Histone H3 [Trimethyl Lys27] Antibody [NBP2-16840] - Mouse brain. Histone H3K27me3 (trimethyl Lys27) antibody diluted at 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27]-Antibody-Immunohistochemistry-Paraffin-NBP2-16840-img0015.jpg)
![Immunohistochemistry-Paraffin: Histone H3 [Trimethyl Lys27] Antibody [NBP2-16840] - Mouse duodenum. Histone H3K27me3 (trimethyl Lys27) antibody diluted at 1:500.Antigen Retrieval: Trilogy™ (EDTA based, pH 8.0) buffer, 15min.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27]-Antibody-Immunohistochemistry-Paraffin-NBP2-16840-img0016.jpg)
![Immunohistochemistry Whole-Mount: Histone H3 [Trimethyl Lys27] Antibody [NBP2-16840] - Paraformaldehyde-fixed 2 day-post-fertilization zebrafish embryo. Histone H3K27me3 (trimethyl Lys27) antibody dilution: 1:100.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27]-Antibody-Immunohistochemistry-Whole-Mount-NBP2-16840-img0023.jpg)
![Immunohistochemistry-Paraffin: Histone H3 [Trimethyl Lys27] Antibody [NBP2-16840] - Mouse intestine. Histone H3K27me3 (Tri-methyl Lys27) stained by Histone H3K27me3 (Tri-methyl Lys27) antibody diluted at 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27]-Antibody-Immunohistochemistry-Paraffin-NBP2-16840-img0025.jpg)
![Dot Blot: Histone H3 [Trimethyl Lys27] Antibody [NBP2-16840] - Various peptide samples were spotted onto positively charged nylon membrane, and blotted with Histone H3 (tri-Methyl Lys27) antibody diluted at 1:2500.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27]-Antibody-Dot-Blot-NBP2-16840-img0012.jpg)
![Western Blot: Histone H3 [Trimethyl Lys27] Antibody [NBP2-16840] - Various whole cell extracts (30 ug) were separated by 15% SDS-PAGE, and the membrane was blotted with Histone H3 antibody (NBP2-16840) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody, and the signal was developed with Trident ECL plus-Enhanced.](http://images.novusbio.com/fullsize/nbp2-16840_rabbit-polyclonal-histone-h3-trimethyl-lys27-antibody-91020242063319.jpg)
![Western Blot: Histone H3 [Trimethyl Lys27] Antibody [NBP2-16840] - Various whole cell extracts (30 ug) were separated by 15% SDS-PAGE, and the membrane was blotted with Histone H3 antibody (NBP2-16840) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.](http://images.novusbio.com/fullsize/nbp2-16840_rabbit-polyclonal-histone-h3-trimethyl-lys27-antibody-9102024201141.jpg)
![Immunocytochemistry/ Immunofluorescence: Histone H3 [Trimethyl Lys27] Antibody [NBP2-16840] - Histone H3 antibody detects Histone H3 protein at nucleus by immunofluorescent analysis.Sample: 293T cells were fixed in 4% paraformaldehyde at RT for 15 min.Green: Histone H3 stained by Histone H3 antibody (NBP2-16840) diluted at 1:500.Red: alpha Tubulin, a cytoskeleton marker, stained by alpha Tubulin antibody [GT114] diluted at 1:1000.](http://images.novusbio.com/fullsize/nbp2-16840_rabbit-polyclonal-histone-h3-trimethyl-lys27-antibody-91020242021980.jpg)
![Western Blot: Histone H3 [Trimethyl Lys27] Antibody [NBP2-16840] - Various whole cell extracts (30 ug) were separated by 15% SDS-PAGE, and the membrane was blotted with Histone H3 antibody (NBP2-16840) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody, and the signal was developed with Trident ECL plus-Enhanced.](http://images.novusbio.com/fullsize/nbp2-16840_rabbit-polyclonal-histone-h3-trimethyl-lys27-antibody-910202420122015.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.](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)
![Immunocytochemistry/Immunofluorescence: Histone H3 [p Ser28, Trimethyl Lys27] Antibody [NB21-1193] - Histone H3 K27me3/pS28 antibody was tested in HeLa cells with DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and DyLight 550 (red).](http://images.novusbio.com/fullsize/Histone-H3-[p-Ser28--Trimethyl-Lys27]-Antibody-Immunocytochemistry-Immunofluorescence-NB21-1193-img0016.jpg "Immunocytochemistry/Immunofluorescence: Histone H3 [p Ser28, Trimethyl Lys27] Antibody [NB21-1193] - Histone H3 K27me3/pS28 antibody was tested in HeLa cells with DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and DyLight 550 (red).")
![Dot Blot: Histone H3 [p Ser28, Trimethyl Lys27] Antibody [NB21-1193] - Analysis of NB21-1193 using the peptides listed above in 100, 10, and 1 picomoles of peptide.](http://images.novusbio.com/fullsize/Histone-H3-[p-Ser28--Trimethyl-Lys27]-Antibody-Dot-Blot-NB21-1193-img0006.jpg)
![Chromatin Immunoprecipitation: Histone H3 [p Ser28, Trimethyl Lys27] Antibody [NB21-1193] - Chromatin from one million formaldehyde cross-linked HeLa cells was used with 2ug of NB21-1193 and 20ul of magnetic IgG beads per immunoprecipitation. A no antibody (No Ab) control was also used. Immunoprecipitated DNA was quantified using quantitative real-time PCR and SYBR green dye, then normalized to the non-precipitated input chromatin, which is equal to one.](http://images.novusbio.com/fullsize/Histone-H3-[p-Ser28--Trimethyl-Lys27]-Antibody-Chromatin-Immunoprecipitation-NB21-1193-img0010.jpg)
![Western Blot: Histone H3 [p Ser28, Trimethyl Lys27] Antibody [NB21-1193] - Tested in (1) HeLa histone prep (2) NIH-3T3 histone prep and (3) C. elegans embryo cell lysate. Observed molecular weight is ~16 kDa.](http://images.novusbio.com/fullsize/Histone-H3-[p-Ser28--Trimethyl-Lys27]-Antibody-Western-Blot-NB21-1193-img0013.jpg)
![Western Blot: Histone H3 [Trimethyl Lys27, p Ser28] Antibody [NBP2-59212] - Cell cycle experiment using the antibody against H3K27me3S28p. WB was performed on cells blocked in different stages of the cell cycle (1=asynchronous population, 2=G1/Sa, 3=G1/Sb, 4=S phase, 5=G2/M, 6=M, 7=G1). The picture shows a nice peak of H3K27me3S28p expression during mitosis. Theoretical molecular weight is ~15 kDa.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27--p-Ser28]-Antibody-Western-Blot-NBP2-59212-img0006.jpg "Western Blot: Histone H3 [Trimethyl Lys27, p Ser28] Antibody [NBP2-59212] - Cell cycle experiment using the antibody against H3K27me3S28p. WB was performed on cells blocked in different stages of the cell cycle (1=asynchronous population, 2=G1/Sa, 3=G1/Sb, 4=S phase, 5=G2/M, 6=M, 7=G1). The picture shows a nice peak of H3K27me3S28p expression during mitosis. Theoretical molecular weight is ~15 kDa.")
![Chromatin Immunoprecipitation: Histone H3 [Trimethyl Lys27, p Ser28] Antibody [NBP2-59212] - ChIP assays were performed using human HeLa cells treated with colcemid, the antibody against H3K27me3S28p and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1 million cells. A titration of the antibody 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 for the promoter of the active gene GAPDH as a negative control, and for the coding regions of the inactive genes MYT1 and TSH2B as positive controls. Figure 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 H3K27me3S28p is preferably present at inactive genes.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27-p-Ser28]-Antibody-Chromatin-Immunoprecipitation-NBP2-59212-img0002.jpg)
![Western Blot: Histone H3 [Trimethyl Lys27, p Ser28] Antibody [NBP2-59212] - Western blot was performed on histone extracts (15 ug, lane 1) from HeLa cells, and on 1 ug of recombinant histone H3 (lane 2) using the antibody against H3K27me3S28p diluted 1:100 in TBS-Tween containing 5% skimmed milk. Observed molecular weight is ~17 kDa.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27--p-Ser28]-Antibody-Western-Blot-NBP2-59212-img0005.jpg)
![ELISA: Histone H3 [Trimethyl Lys27, p Ser28] Antibody [NBP2-59212] - To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody directed against H3K27me3S28p. 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 1:4,300.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27--p-Ser28]-Antibody-ELISA-NBP2-59212-img0001.jpg)
![Dot Blot: Histone H3 [Trimethyl Lys27, p Ser28] Antibody [NBP2-59212] - A Dot Blot analysis was performed to test the cross reactivity of the antibody against H3K27me3S28p with peptides containing other modifications of histone H3 and H4 and the unmodified H3K27S28 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 1:2,000. The figure shows a high specificity of the antibody for the modification of interest.](http://images.novusbio.com/fullsize/Histone-H3-[Trimethyl-Lys27--p-Ser28]-Antibody-Dot-Blot-NBP2-59212-img0004.jpg)
![Western Blot: Histone H3 [Trimethyl Lys27] Antibody (C.15200181) [NBP2-59295] - Histone extracts (15 ug) from HeLa cells were analysed by Western blot using the antibody against H3K27me3 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-Lys27-Antibody-C-15200181-Western-Blot-NBP2-59295-img0001.jpg "Western Blot: Histone H3 [Trimethyl Lys27] Antibody (C.15200181) [NBP2-59295] - Histone extracts (15 ug) from HeLa cells were analysed by Western blot using the antibody against H3K27me3 diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. Observed molecular weight is ~17 kDa.")
![Immunocytochemistry/Immunofluorescence: Histone H3 [Trimethyl Lys27] Antibody (C.15200181) [NBP2-59295] - HeLa cells were stained with the antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa Fluor 594. 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-C-15200181-Immunofluorescence-NBP2-59295-img0004.jpg)
![Chromatin Immunoprecipitation: Histone H3 [Trimethyl Lys27] Antibody (C.15200181) [NBP2-59295] - ChIP assays were performed using human HeLa cells, the antibody against H3K27me3 and optimized PCR primer sets for qPCR. ChIP was performed with the AutoHistone ChIP-seq kit on sheared chromatin from 1 million cells. A titration of the antibody 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 for the promoters of the active genes c-fos and GAPDH as negative controls, and for the coding regions of the inactive genes MYT1 and TSH2B as positive controls. Figure 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 H3K27me3 is preferably present at inactive genes.](http://images.novusbio.com/fullsize/Histone-H3-Trimethyl-Lys27-Antibody-C-15200181-Chromatin-Immunoprecipitation-NBP2-59295-img0003.jpg)
![ELISA: Histone H3 [Trimethyl Lys27] Antibody (C.15200181) [NBP2-59295] - Cross reactivity of the antibody directed against H3K27me3. To test the specificity an ELISA was performed using a serial dilution of the antibody against H3K27me3. The wells were coated with peptides containing the unmodified H3K27 region as well as the mono-, di- and trimethylated H3K27 and the trimethylated H3K9. Figure shows a high specificity of the antibody for the peptide containing the modification of interest.](http://images.novusbio.com/fullsize/Histone-H3-Trimethyl-Lys27-Antibody-C-15200181-ELISA-NBP2-59295-img0002.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 [Trimethyl Lys27] Antibody (BT164) - Chimeric [NBP3-12020] - Nuclear lysate of HeLa(A) (0.0003ug/ml), Jurkat(B) (0.0003ug/ml) and K562(C) (0.01ug/ml) (35ug protein in RIPA buffer) were resolved on a SDS PAGE gel and blots were probed with the chimeric rabbit version of BT164 (NBP3-12020) before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.](http://images.novusbio.com/fullsize/Histone-H3-Trimethyl-Lys27-Antibody-BT164-Chimeric-Western-Blot-NBP3-12020-img0001.jpg "Western Blot: Histone H3 [Trimethyl Lys27] Antibody (BT164) - Chimeric [NBP3-12020] - Nuclear lysate of HeLa(A) (0.0003ug/ml), Jurkat(B) (0.0003ug/ml) and K562(C) (0.01ug/ml) (35ug protein in RIPA buffer) were resolved on a SDS PAGE gel and blots were probed with the chimeric rabbit version of BT164 (NBP3-12020) before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.")
![Immunocytochemistry/Immunofluorescence: Histone H3 [Trimethyl Lys27] Antibody (BT164) - Chimeric [NBP3-12020] - Immunofluorescence analysis of paraformaldehyde fixed HeLa cells on Shi-fix(TM) coverslips stained with the chimeric rabbit IgG version of BT164 (NBP3-12020) at 10 ug/ml for 1h followed by Alexa Fluor(R) 488 secondary antibody (2 ug/ml), showing membrane staining. The nuclear stain is DAPI (blue) and the actin stain is phalloidin (red). Panels show from left-right, top-bottom NBP3-12020, DAPI, merged channels and an isotype control. The isotype control was an unknown specificity antibody followed by staining with Alexa Fluor(R) 488 secondary antibody.](http://images.novusbio.com/fullsize/Histone-H3-Trimethyl-Lys27-Antibody-BT164-Chimeric-Immunocytochemistry-Immunofluorescence-NBP3-12020-img0002.jpg)