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).