Rabbit IgG TrueBlot is provided as 1000X solution. We recommend incubating the blots from minigels in sealed bags with minimal volume (2-3 mLs). If used conservatively at 2.5mls/blot will yield enough reagent for 20 blots. Rabbit IgG TrueBlot HRP-conjugated antibody reacting with goat IgGs for optimal signal detection in IP/WB experiments. Specific conditions for each protein and antibody combination should be specifically optimized by the end user. Note that there are three key procedural considerations: 1. Protein A or G should not be used for the IP. Use of protein A or G beads with the rabbit TrueBlot will result in contaminating bands. For IP, Anti-Rat IgG beads, or Anti-Rabbit IgG beads should be used for rat or rabbit immunoprecipitating antibodies, respectively. 2. Immunoprecipitate should be completely reduced. 3. BLOTTO/Milk should be used as the blocking protein for the immunoblot.