Horseradish peroxidase hrp conjugated anti mouse and anti rabbit igg antibodies

Tunicamycin was purchased from Wako Pure Chemical Industries (Osaka, Japan). 2-Deoxy-D-glucose and rabbit polyclonal antibodies against glial fibrillary acidic protein (GFAP), doublecortin (DCX), and HRD1 (C-terminal) were purchased from Sigma-Aldrich (St. Louis, MO). Mouse monoclonal antibodies against nestin, βIII-tubulin, neuronal nuclei protein (NeuN; A60), and microtubule-associated protein-2 (MAP-2) were purchased from Millipore (Temecula, CA). A mouse monoclonal antibody against KDEL (10C3) was purchased from Stressgen Biotechnologies (Victoria, British Columbia, Canada). A rabbit polyclonal antibody against cleaved Notch1 (Val1744) was purchased from Cell Signaling Technology (Danvers, MA). A goat polyclonal antibody against Notch3 (M-20) and a rabbit polyclonal antibody against GADD 153 (CHOP) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Horseradish peroxidase (HRP)-conjugated anti-mouse and anti-rabbit IgG antibodies were purchased from GE Healthcare (Buckinghamshire, United Kingdom). An Alexa Fluor 488-conjugated anti-mouse IgG antibody and an Alexa Fluor 546-conjugated anti-rabbit IgG antibody were purchased from Life Technologies (Grand Island, NY). Western Lightning Chemiluminescence Reagent Plus was obtained from PerkinElmer Life Science (Waltham, MA).

RPMI 1640 and DMEM were obtained from Sigma-Aldrich, St. Louis, MO. Antibodies against p-PYK2 (sc-293142), p-Vav (sc-16408-R), and Vav (sc-17831) were obtained from Santa Cruz Biotechnology Inc., Dallas, TX. Anti-phosphotyrosine (11-263-C100) and anti-Syk (11-376-C100) antibodies were purchased from Exbio, Vestec, Czech Republic. Anti-Pyk2 antibody (P3902) was purchased from Sigma-Aldrich, St. Louis, MO. Antibody against p-Syk (MAB6459) was purchased from R&D systems, Minneapolis, MN. Rabbit polyclonal anti-cAMP antibody for competitive ELISA was obtained from GenScript, Piscataway, NJ. Horseradish peroxidase (HRP)-conjugated anti-mouse and anti-rabbit IgG antibodies were purchased from GE Healthcare, Piscataway, NJ. HBSS buffer (10 mM HEPES, pH 7.4, 140 mM NaCl, 5 mM KCl) complemented with 2 mM CaCl₂, 2 mM MgCl₂, 1% (w/v) glucose, and 1% (v/v) fetal calf serum (FCS)) was prepared for flow cytometry experiments. Zymosan A was purchased from Sigma-Aldrich, St. Louis, MO. For labeling, Zymosan At 10 mg/ml was incubated with 50 ug/ml of fluorescein isothiocyanate (FITC; Sigma-Aldrich, St. Louis, MO) in 300 mM sodium bicarbonate buffer (pH 9.2) with overnight rotation at 4 °C. Unbound FITC was removed by extensive washing with HBSS buffer, and FITC labeled zymosan (FITC-zymosan) was stored at −20 °C.