A Convenient Western-blot like Protocol
The AbSurance™ Histone Antibody Specificity Arrays are essential screening tools to evaluate the specificity and cross-reactivity of antibodies against histones H2A, H2B, H3, H4, and their post-translational modifications. Two arrays provide a total of 89 unique high quality peptides in 10 ng and 100 ng quantities to easily detect both strong and weak cross-reactivity. The H3 Array provides 8 acetylated, 27 methylated, 5 phosphorylated, and 6 unmodified peptides of histone H3. The H2A, H2B, H4 Array offers 13 acetylated, 16 methylated, 6 phosphorylated, and 8 unmodified peptides of H2 and H4. Additional rat, sheep, rabbit, and mouse primary antibodies are spotted on each array for convenient built-in positive controls. The arrays are designed on PVDF membranes for a simple Western blot-like assay; no need for additional equipment or software. Detection is rapid with standard chemiluminescent reagents, and spot identification is as easy as aligning your array with the alignment grid provided.
AbSurance detects specific and non-specific interactions.
Detection of specific interactions using EMD Millipore’s antibody to trimethyl histone H3 lysine 4 (Catalogue No. 05-745R). AbSurance™ array shows expected specificity for H3K4me3 peptide (9A and 9B, green box) without cross-reaction to other peptides including unmodified H3, H3K4 (6A and 6B), H3K4me (7A and 7B), and H3K4me2 (8A and 8B). Primary antibody used at 1:2,000 dilution. Positive control rabbit IgG shown at position 12H (blue box).
Detection of non-specific interactions using an antibody to acetyl histone H3 lysine 56 obtained from Supplier E. AbSurance™ array data demonstrate weak recognition of the target H3K56ac peptide (6G and 6H, green box) but strong cross-reactivity with H3K9ac (1C and 1D, red box). Primary antibody used at 1:2,000 dilution. Positive control rabbit IgG shown at position 12H (blue box).