Hrp conjugated sheep anti mouse ig ab

The following were purchased: PIPES, bovine serum albumin (BSA), EGTA, EDTA, D-glucose, NaF, Na4P2O7, Na3VO4, 2-ME, NP-40, FMLP (Sigma, St. Louis, MO); crystallized human serum albumin (HSA) (Miles Laboratories, Elkhart, IN); fetal calf serum (FCS) and RPMI 1640 containing 25 mM HEPES and L-glutamine (BioWhittaker, Walkersville, MD); Percoll, (Pharmacia, Piscataway, NJ); anti-SYK mAb, 4D10 (Santa Cruz Biotechnology, Santa Cruz, CA); a penicillin (BPO)-specific IgE was partially purified from the sera of penicillin allergic patients as previously described [7 (link)]; BPO2 (benzylpenicilloyl-di-octamine) was synthesized and purified as described previously [10 (link),11 (link)]; monoclonal anti-human IgE Ab (6061P) (Hybridoma Labs, Baltimore, MD); antiHRP-conjugated donkey anti-rabbit Ig Ab, HRP-conjugated Sheep anti-mouse Ig Ab, protein G sepharose beads (Amersham Life Science, Arlington Heights, IL); monoclonals anti-FceRI antibodies 22E7 and 15A5 (gift of former Hoffman-LaRoche)(measured total FceRI and unoccupied FceRI, respectively). Countercurrent elutriation and labeling with antibodies for flow cytometry was conducted in PAG containing 0.25 % BSA in place of 0.003 % HSA. ESB is Novex electrophoresis sample buffer containing 5 % 2-mercaptoethanol. SDS stripping buffer for Western blots was 65 mM Tris (pH 6.7), 100 mM 2-mercaptoethanol and 2% SDS.