How should blocking peptides be used for western blotting?

For western blotting, we recommend mixing the primary antibody in the appropriate dilution buffer (see product datasheet) with at least 1.0 ug of peptide per 1.0 ul of primary antibody and incubating for 20 minutes with agitation before adding the mixture to the membrane. For example, in 10 ml of primary antibody dilution buffer with 10 ul antibody (1:1000 final dilution), you would add 10 ul peptide stock that is at 1.0 mg/ml (1:1000 final dilution). After adding to the membrane, the antibody/peptide mixture should incubate overnight at 4C with agitation followed by incubation with the secondary antibody as usual.

Last updated: September 12, 2024