Histone H3 [Trimethyl Lys27] Antibody - BSA Free

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).

ELISA: Histone H3 [Trimethyl Lys27] Antibody [NBP2-54614]

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.

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.