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![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)