IP and WB Kit for FLAGr Epitope Tag 1Kit

Mouse IgG TrueBlot® ULTRA 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). Mouse IgG TrueBlot® ULTRA is a HRP-conjugated monoclonal secondary antibody reacting with mouse IgGs for optimal signal detection in immunoprecipitation/immunoblot 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. TrueBlot® IP and WB Kit for DYKDDDDK (FLAG®) Epitope Tag 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. Anti-Mouse Ig IP Beads: 2.5 mL Binds 0.4 mg Ig/mL beads; 8. Antibody for the detection of FLAG™ conjugated proteins (MOUSE) Monoclonal Antibody: 100 µg; 9. 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.