Western blotting, cyclin-dependent kinases. 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/immunoblot applications. Terms to be familiar with before you start to solve the test: immunoprecipitation, affinity chromatography. TrueBlot preferentially detects the non-reduced form of mouse IgG over the reduced, SDS-denatured form of IgG. We also take advantage of RNAi or chemical inhibitors to modulate OGT and OGA activities before HCF-1 immunoprecipitation. Antigens isolated by IP are analyzed by SDS-PAGE or Western blotting. Mouse IgG TrueBlot ULTRA is ideal for use in protocols involving immunoblotting of immunoprecipitated proteins. By using the host-cell factor 1 (HCF-1), a known O-GlcNAcylated protein, we immunoprecipitated HCF-1 from transfected HEK293T cells or endogenous HCF-1 from HeLa cells to detect its O-GlcNAc levels by Western blotting. isolate a specific antigen from a mixture, using the antigen-antibody interaction. It is easy to generate publication-quality IP/WB data with Mouse IgG TrueBlot® ULTRA. Use it in place of your conventional HRP Anti-Mouse IgG immunoblotting secondary antibody. Mouse IgG TrueBlot® ULTRA is the unique horseradish peroxidase conjugated Anti-Mouse IgG imonoclonal secondary antibody which enables detection of immunoblotted target protein bands, without hindrance by interfering immunoglobulin heavy and light chains from your IP antibody. The TrueBlot® Immunoprecipitation and Western Blot Kit for DYKDDDDK (FLAG®) Epitope Tag contains the critical supporting reagents, buffers, and substrates for immunoprecipitation and Western blotting of samples containing the DYKDDDDK (FLAG®) Epitope Tag using TrueBlot monoclonal secondary antibody in conjunction with Rockland's Antibody for the detection of FLAG™ conjugated proteins (MOUSE) Monoclonal Antibody (see application_notes). The Anti-DYKDDDDK (FLAG®) antibody included in this Western blot kit detects FLAG®-tagged recombinant proteins fused with FLAG® tag either at the amino-terminus or carboxy-terminus of the protein. The hydrophilic character of this sequence increases the likelihood that it will be located on the surface of FLAG®-tagged recombinant proteins for reaction with anti-DYKDDDDK (FLAG®) antibodies. The FLAG® epitope is a small but highly immunogenic peptide DYKDDDDK (N-Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys-C) which allows fusion proteins to retain their original conformation and function.