Extended Data Figure 3: Quality control of the H2A Q105me antibody.
From: Glutamine methylation in histone H2A is an RNA-polymerase-I-dedicated modification
a, Examples of dot blots of dilution series (1:2) of unmodified or methylated peptide spotted on 0.22 μm PVDF membrane and developed using the anti-H2A Q105me antibody. b, Left, western blot on lysates of cells harbouring either WT or Q105A H2A as sole source of histones; right, human lysate from MCF10A cells. All western blots were probed with the anti-H2AQ105me-specific antibody or anti-H2A upon stripping, respectively. c, Peptide competition by western blotting with the indicated peptides shows that only the methylated peptide can compete the signal of MCF10A lysates (peptide concentration was at 1 μg ml−1). d, Direct enzyme-linked immunosorbent assay (ELISA) against the indicated peptides. Biotinylated peptides were immobilized on streptavidine-coated 96-well plates and the ELISA was performed using a 1:20,000 dilution of the anti-Q105me antibody. The ELISA was developed using horseradish peroxidase (HRP)-coupled secondary anti-rabbit antibody and 3,3′, 5,5′′-tetramethylbenzidine (TMB) as substrate.
