Goat TrueBlot® Set (IP Agarose beads)

• Related Content: TrueBlot monoclonal secondary antibodies for IP-WB
  

Goat IgG TrueBlot® is a unique horseradish peroxidase conjugated Anti-Goat IgG monoclonal secondary antibody. Goat IgG TrueBlot® enables detection of immunoblotted target protein bands, without hindrance by interfering immunoprecipitating immunoglobulin heavy and light chains. It is easy to generate publication-quality IP/Western Blot data with Goat IgG TrueBlot®, simply substitute the conventional HRP Anti-Goat IgG blotting reagent with Goat IgG TrueBlot® and follow the prescribed protocol for sample preparation and immunoblotting. Goat IgG TrueBlot® is ideal for use in protocols involving immunoblotting of immunoprecipitated proteins. TrueBlot preferentially detects the non-reduced form of goat IgG over the reduced, SDS-denatured form of IgG. When the immunoprecipitate is fully reduced immediately prior to SDS-gel electrophoresis, reactivity of Goat IgG TrueBlot® 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. Goat IgG TrueBlot may also be used for detection in immunoblotting assays that do not employ immunoprecipitation. Goat IgG TrueBlot® is provided as 1000X solution. To conserve reagent, we recommend incubating the blots from minigels in sealed bags (removing as much air as possible) with minimal volume (2-3 mLs). If used conservatively at 2.5mls/blot will yield enough reagent for 20 blots. Note that there are three key procedural considerations: 1. Protein A or G beads may be used with the mouse, goat and sheep TrueBlot secondaries, but not with the rabbit TrueBlot secondary. Use of protein A or G beads with the rabbit TrueBlot will result in contaminating bands. 2. Immunoprecipitate should be completely reduced. 3. BLOTTO/Milk should be used as the blocking protein for the immunoblot. MB-70 or BSA is not an effective blocker. Goat TrueBlot Set Components: 1. Goat IgG TrueBlot® HRP-conjugated monoclonal secondary antibody reacting with goat IgGs for optimal signal detection in immunoprecipitation/immunoblotting experiments. 2. Anti-Goat Ig IP Beads: 2.5 mL. Binds 1 mg Ig/mL beads. 3. Western blot incubation tray. Special Notes: Upon initial use of the IP beads, we recommend that the vial be inverted several times to get the beads into suspension. We recommend using a large bore pipet to pipet up the liquid for use. For storage of the opened vial of beads, we recommend that the vial cap be sealed with parafilm to help prevent evaporation of the buffer. 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.