Rat Pure-Blot anti-Mouse IgG (H+L) Secondary Antibody (eB144) [Biotin]

This antibody is ideal for use in protocols involving immunoblotting of immunoprecipitated proteins. This antibody preferentially detects the non-reduced form of mouse IgG (IgG1, IgG2a, IgG2b, IgG3) over the reduced, SDS-denatured form of IgG. To achieve best results when detecting mouse IgG1 subtypes, we recommend performing a dot blot or titration to determine the optimal dilution factor for your desired application. When the immunoprecipitate is fully reduced immediately prior to SDS-gel electrophoresis, reactivity of This antibody with the 55 kDa heavy chains and the 23 kDa light chains of the immunoprecipitating antibody is minimized thereby eliminating interference by the heavy and light chains of the immunoprecipitating antibody in IP/Western blot applications. Applications include studies examining post-translational modification (e.g., phosphorylation or acetylation) or protein-protein interactions. Note: There are two key procedural considerations: 1. When using any Pure-Blot reagent, ensure there is complete reduction of the lysate. 2. Use BLOTTO/milk powder for complete and effective blocking of the western blot. All recommended dilutions for listed applications are intended as an initial recommendation, specific conditions for each protein and antibody combination should be specifically optimized by the end user.