This TrueBlot(R) Immunoprecipitation and Western Blot Kit for 6X HIS Epitope Tag allows for the detection of 6X HIS-tagged recombinant protein present in cell lysates provided by the user. Mouse TrueBlot(R) ULTRA Antibody Peroxidase Conjugate was prepared from tissue culture supernatant by Protein G affinity chromatography. Assay by Immunoelectrophoresis resulted in a single precipitin arc against anti-Mouse Serum. Reactivity is observed against native Mouse IgG by both Western blot and ELISA. TrueBlot(R) Immunoprecipitation and Western Blot Kit for 6X HIS Epitope Tag allows for the immunoprecipitation and detection of 6X HIS-tagged recombinant proteins present in cell lysates provided by the user. After protein separation by SDS-PAGE and transfer, the membrane is probed with monoclonal Anti-6X HIS. Detection of the membrane bound antibody-antigen complex is achieved by the addition of Mouse IgG TrueBlot(R) ULTRA HRP. The TrueBlot(R) Immunoprecipitation and Western Blot Kit for 6X HIS Epitope Tag contains the critical supporting reagents, buffers, and substrates for immunoprecipitation and Western blotting of samples containing a 6X HIS Epitope Tag using TrueBlot second step immunoblotting reagents in conjunction with Rockland's Anti-6X HIS (MOUSE) Monoclonal Antibody. TrueBlot technology enables unhindered detection of protein bands of interest which would otherwise be obscured by the presence of reduced and denatured heavy and light chain immunoglobulin in the blot (as detected by the conventional immunoblotting HRP anti-mouse IgG reagent). Mouse IgG TrueBlot(R) ULTRA is the unique horseradish peroxidase conjugated anti-mouse IgG immunoblotting second step reagent which enables detection of immunoblotted target protein bands, without hindrance from interfering immunoglobulin heavy and light chains from your IP antibody. Use it in place of your usual HRP anti-mouse IgG immunoblotting second step reagent. It is easy to generate publication-quality IP/WB data with Mouse IgG TrueBlot(R) ULTRA. Mouse IgG TrueBlot ULTRA is ideal for use in protocols involving immunoblotting of immunoprecipitated proteins. TrueBlot preferentially detects the non-reduced form of mouse IgG over the reduced, SDS-denatured form of IgG. When the immunoprecipitate is fully reduced immediately prior to SDS-gel electrophoresis, reactivity of Mouse IgG TrueBlot ULTRA 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/immunoblotting applications. Applications include studies examining post-translational modification (e.g., phosphorylation or acetylation) or protein-protein interactions.
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