Confocal immunofluorescent analysis of HeLa cells using Di-Methyl-Histone H3 (Lys9) mAb(green). Actin filaments have been labeled with DY-554 phalloidin (red).

Di-Methyl Histone H3 (Lys9) mAb specificity was determined by peptide ELISA. The graph depicts the binding of the antibody to pre-coated di-methyl histone H3 (Lys9) peptide in the presence of increasing concentrations of various competitor peptides. As shown, only the di-methyl histone H3 (Lys9) peptide competed away binding of the antibody.

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4x10e6 HeLa cells and either 20ul of H5110-14D2 or 2ul of Normal Rabbit IgG using SimpleChIP Enzymatic Chromatin IP Kit (Magnetic Beads). The enriched DNA was quantified by Real-Time PCR using primers specific for the transcriptionally active GAPDH and RPL30 genes, the inactive AFM gene, and the heterochromatic alpha Satellite repeat element. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

Flow cytometric analysis of HeLa cells using Di-Methyl-Histone H3 (Lys9) (H5110-14D2) (blue) compared to concentration-matched Rabbit (DA1E) mAb IgG XP Isotype Control (red). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor 488 Conjugate) was used as a secondary antibody.