Mouse IgG TrueBlot® ULTRA is provided as 1000X solution. 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. 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. 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. Mouse IgG TrueBlot® ULTRA HRP-conjugated secondary antibody reacting with mouse IgGs for optimal signal detection in immunoprecipitation/immunoblotting experiments. 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. A sample of Anti-Alpha-Tubulin (MOUSE) Monoclonal Antibody Cat #(200-301-880S), has been provided to be used as a loading control at the user’s discretion. Dilutions are to be optimized by the user. Mouse TrueBlot® Western Blot Kit Components: 1. Mouse IgG TrueBlot ULTRA: 50 µL 18-8817-31, 2. TrueBlot Enhancer Solution: 25 mL; 3. TrueBlot Blocker: 10 g; 4. TrueBlot Assay Buffer: 30 mL, 20X; 5. TrueBlot Substrate A: 12.5 mL; 6. TrueBlot Substrate B: 12.5 mL; 7. Western Blot Incubation Tray. 8. Anti-Alpha-Tubulin (MOUSE) Monoclonal Antibody - 200-301-880S.