Sephadex g 75

Mature dry seeds of B. monosperma were collected from Dr. R.M.L. Avadh University Campus, Faizabad (Uttar Pradesh, India). The plant was authenticated by plant taxonomist Dr. Tariq Husain, National Botanical Research Institute, Lucknow. The voucher specimen (specimen accession number 98180-2011) was deposited at the NBRI herbarium. N-tosyl-L-phenylalanine chloromethyl ketone (TPCK) treated bovine trypsin, synthetic substrate N-α-benzoyl-DL-arginine-p-nitroanilide (BApNA), standard inhibitor phenyl methyl sulphonyl fluoride (PMSF), acrylamide, and other electrophoretic reagents were purchased from Sigma Chemical Co. (St. Louis, Mo, USA). Sephadex G-75 was purchased from Amersham Biosciences (Uppsala, Sweden). Trypsin-Sepharose CL-4B, azocasein, and protein molecular weight markers were purchased from Sisco Research Limited (Mumbai, India). All other chemicals used were of analytical grade. H. armigera were from a laboratory colony which was obtained from Narendra-Dev University of Agriculture and Technology, Kumarganj, Faizabad, India. Insects were housed at 28 ± 2°C, 60% relative humidity with a photoperiod of 14 h light and 10 h dark.

DTT, SBTI, and EAM were obtained from Merck (Beijing, China). Fibrin, Fibrinogen, pNA, PMSF, DMSO, EGTA, PHMB, leupeptin, and pepstatin A were procured from Sigma-Aldrich (MO, USA). Chromogenic synthetic substrates such as D-Val-Leu-Arg-pNA (V6258), D-Val-Leu-Lys-pNA (V7127), D-Phe-Pip-Arg-pNA (P7027), and N-Succ-Ala-Ala-Pro-Phe-pNA (S7388) were also purchased from Sigma-Aldrich. HT29 human adenocarcinoma cell line was purchased from Merck (Darmstadt, Germany). Phenyl-Sepharose 6FF, DEAE-Sepharose CL-6B, Sephadex G-75, and marker proteins were purchased from Amersham Biosciences (Sweden). The rest of the chemicals were of analytical grade from resident providers.

N-α-Benzoyl-DL-arginine-p-nitroanilide (BApNA), soybean trypsin inhibitor (SBTI), Bovine serum albumin (BSA), acrylamide, bis-acrylamide, and other electrophoretic reagents were procured from Sigma Chemical Co. (St. Louis, Mo, USA). Sephadex G-75 was purchased from Amersham Biosciences (Uppsala, Sweden). Trypsin-Sepharose CL-4B, low molecular weight markers and azocasein were purchased from Sisco Research Limited (Mumbai, India). All other chemicals and reagents used were of analytical grade.