Non-specific binding of an antibody to proteins other than the antigen can sometimes occur. This is usually more common with polyclonal antibodies, but can also occur with monoclonals as well. To determine which band or staining is specific, the AKT immunizing peptide blocking experiment can be performed. Prior to the staining protocol, the anti-AKT antibody is neutralized (incubated with an excess of peptide that corresponds to the epitope recognized by the antibody). The AKT antibody that is bound to the blocking peptide is no longer available to bind to the epitope present in the protein on the Western blot or in the cell. The neutralized antibody is then used side-by-side with the antibody alone, and the results are compared. By comparing the staining from the blocked antibody versus the antibody alone, you can see which staining is specific: this staining will be absent from the Western blot or immunostaining performed with the neutralized antibody.
Category
Proteins, Peptides and Small Molecules
Product Group
Peptides
Conjugate
Unconjugated
Purity
>95%
Formulation
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
