2 4 6 trinitrobenzenesulphonic acid tnbs

Phosphate-buffered saline (PBS) was from Avantor Performance Materials Poland S.A. (Gliwice, Poland). Arachidonate, collagen, and ADP were from Chrono-Log Corp. (Havertown, PA, USA). Dimethyl sulfoxide, cytochrome c, sodium dodecyl sulphate (SDS), Ellman's reagent (5,5′-dithiobis-2-nitrobenzoic acid, DTNB), glutathione (reduced), HCl, 2,4,6-trinitrobenzenesulphonic acid (TNBS), ethanol, ethyl acetate, guanidine hydrochloride, xylenol orange, Fe(NH₄)₂(SO₄)₂, and perchloric acid were from Sigma-Aldrich (St. Louis, MO, USA). Pierce™ BCA Protein Assay Kit was from Thermo Fisher Scientific (Waltham, Massachusetts, USA). Fluorolabelled monoclonal antibodies (moAbs)—anti-CD61/PerCP, antiCD62/PE, PAC-1/FITC, isotype antibodies, and CellFix were from Becton Dickinson (San Diego, CA, USA).

Methylglyoxal (MGO), Protein-A-agarose pre-packed affinity column, Congo red (CR), Thioflavin T (ThT), 8-anilinonaphthalene-1-sulfonic acid (ANS), sodium dodecyl sulphate, dialysis tubing, 2,4,6-trinitrobenzene sulphonic acid (TNBS) and standard molecular weight marker were purchased from Sigma Chemical Company (St. Louis, MO, USA). D-glucose was purchased from Qualigens, India. Nitroblue tetrazolium (NBT) dye, 2,4-dinitrophenylhydrazine (DNPH) and silver nitrate were obtained from SRL, India. Acrylamide, bisAcrylamide, ammonium persulphate and N,N,N’,N’-tetramethylethylene- diamine (TEMED) were purchased from Bio-Rad Laboratories, USA. All other reagents were of highest analytical grade available. The study protocol was approved by the Institutional Ethics Committee (IEC), Faculty of Medicine, Aligarh Muslim University, India. Furthermore, the blood sample was voluntarily donated by MAK and the same was used for IgG isolation. This was reported to the IEC.

Alcalase (EC 3.4.21.62) (2.4 unit/g) from Bacillus licheniformis and papain from papaya (Carica papaya) latex (E.C.3.4.22.2) (3 units/g) were obtained from Siam Victory Chemicals Co, Ltd. (Bangkok, Thailand). 2,4,6-trinitrobenzenesulphonic acid (TNBS) was procured from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). All chemicals were of analytical grade.