Monoclonal anti phosphotyrosine antibody

Calcium ionophore (A23187), benzamidine hydrochloride, N-acetyl-Leu-Glu-His-Asp trifluoro methylcoumarin (AC-LEHD-FMC), acetyl-Asp-Glu-Val-Asp-7-amido-4-methylcoumarin (AC-DEVD-AMC), sodium orthovanadate, dimethyl sulfoxide (DMSO), fluorescein isothiocyanate (FITC)-labeled annexin V, 5-(and-6)-chloromethyl-2′,7′-dichlorodihydrofluorescein diacetate acetylester (CM-H2DCFDA), CHAPS, rhodamine 123, leupeptin hydrochoride, N-(2-Hydroxyethyl) piperazine-N′-ethanesulfonic acid (HEPES), fura-2/AM, monoclonal anti-phosphotyrosine antibody, acridine orange 10-nonyl bromide (NAO) and dithiothreitol (DTT) were from Sigma Chemicals, St. Louis (USA). Monoclonal anti-cytochrome c antibody and anti-β-actin were from Epitomics Burlingame, CA (USA). Anti-Caspase-3 antibody was from Santa Cruz Biotechnology, Inc. Texas (USA). Collagen type-I was from Chrono-log Corporation, Pennsylvania (USA). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and 1, 1-diphenyl-2-picrylhdrazyl (DPPH) were from HiMedia Laboratories, Mumbai (India). Lactate dehydrogenase (LDH) kit was from AGAPPE diagnostics Ltd., Kerala (India). γ-glutamyl p-nitroanilide and glycylglycine were from Sisco Research laboratories Pvt Ltd., Mumbai (India). All other reagents were of analytical grade.

Monoclonal anti-polyHistidine-peroxidase antibody produced in mouse was purchased from Sigma (Cat. No. A7058). Monoclonal anti-phosphotyrosine antibody produced in mouse was purchased from Sigma (Cat. No. P4110). HRP conjugated anti-mouse IgG (H + L) antibody was purchased from Promega and used as the secondary antibody for detecting phosphotyrosine (Cat. No.W402B). AP-conjugated goat anti-rabbit IgG was purchased from Cedarlane (Cat. No. CLCC43008). Peroxidase-conjugated goat anti-mouse IgG was purchased from Cedarlane (Cat. No. 115-036-003). Penta-his antibody was purchased from Qiagen (Cat. No. 34660)