5 bromo 4 chloro 3 indolyl phosphate bcip nitroblue tetrazolium nbt

Mentha arvensis leaves were collected from the state of Assam, India. Silver nitrate (AgNO₃), poly(ethylene glycol) (PEG) and 5-bromo-4-chloro-3-indolyl phosphate (BCIP)/nitro blue tetrazolium (NBT) were purchased from Merck (India). Dulbecco’s Modified Eagle’s Medium (DMEM), Roswell Park Memorial Institute (RPMI)-1640 culture media, fetal bovine serum (FBS), penicillin–streptomycin antibiotic solution, phytohemagglutinin (PHA), 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and radio-immunoprecipitation assay (RIPA) buffer were purchased from HiMedia (Mumbai, India). Heparin, ethidium bromide (EtBr) and trypan blue were purchased from SRL (India); poly-l-lysine, Histopaque, bisbenzimide (Hoechst 33342), paraformaldehyde, acridine orange, propidium iodide (PI), RNase, NaBH₄ and anti-human primary antibodies used were procured from Sigma-Aldrich (St Louis, MO, USA). ALP-linked goat anti-rabbit secondary antibodies were purchased from Abcam (UK).

Primary mouse monoclonal antibodies against influenza A nucleoprotein and influenza B nucleoprotein, secondary anti-mouse monoclonal with and without alkaline phosphatase enzyme (ALP) conjugation antibodies, and all respiratory virus protein antigens including recombinant influenza A nucleoprotein, influenza B virus-infected Madin–Darby Canine Kidney (MDCK) Cell lysate, RSV, parainfluenza virus, and adenovirus proteins were supplied from Fitzgerald, MA, USA, East Coast Bio, TN, USA, and Meridian Life Sciences, TN, USA, and Seracare (MA, USA). Forty-percent acrylamide/bis solution (29 : 1) was supplied from Bio-Rad Laboratories Inc. (CA, USA). Reagents used in sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) and western blotting analyses including 5-bromo-4-chloro-3-indolyl phosphate (BCIP)/nitro blue tetrazolium (NBT), and device components were purchased from Merck Millipore (MA, USA). Bovine serum albumin (BSA), ammonium sulfate, β-mercaptoethanol, and coomassie brillant blue R250 solution were supplied from Sigma-Aldrich (MO, USA). All other chemicals and tested proteins were of analytical grade were supplied from Sigma-Aldrich (MO, USA), unless stated otherwise. Ultrapure water (18.25 MΩ cm) were obtained using Milli-Q system (Merck Millipore) and used throughout the experiments.