Measuring Histone Acetylation
We extracted proteins from tissues using the EpiQuick Total Histone Extraction Kit (EpiGentek, Farmingdale, NY). We then stored the extracts at -80°C overnight, and measured H3 histone acetylation in triplicate the next day using the Total Histone H3 Acetylation Detection Fast Kit (EpiGentek, Farmingdale, NY). Briefly, the Total Histone H3 Acetylation Detection Fast Kit detects H3 acetylation by ELISA. Strip wells are coated with an anti-acetyl histone H3 antibody that captures acetyl histone H3 from the protein extract and allows the experimenter to colorimetrically detect if histone H3 is acetylated, and colorimetrically quantify the amount of the acetylated histone H3. The H3 histone presence is detected and measured using a labeled detection antibody and a color development reagent. We measured the resultant color change with a plate reader (Bio Tek, Synergy HTX Multimode Reader, Agilent, Santa Clara CA. USA).