Reagents, proteins and conjugates
Fluorescein isothiocyanate (FITC), methyl α-D-mannopyranoside (MαM), methyl α-D-galactoside (MαG), cellobiose ( 4-O-β-D-Glucopyranosyl-D-glucose), orthophenylene diamine (OPD), horse radish peroxidase (HRP), soybean trypsin inhibitor, galactose, Tween-20, Coomassie brilliant blue (G-250 and R-250), soluble guar galactomannan, ADP, fibrinogen, and rabbit antibodies to human albumin were purchased from Sigma Aldrich, Bangalore, India. Fluorescent-labeled amyloid β peptide Aβ-42 monomer (F-Aβ; HiLyte Fluor-488-Aβ) was purchased from Anaspec, California. Polystyrene 96 well microplates (Maxisorb BREAKAPART and Polysorb BREAKAPART) were purchased from Nunc, Denmark and antibodies to human IgA, IgG and IgM raised in rabbit, from Dako, Denmark.
Jacalin was prepared from jack fruit (Artocarpus integrifolia ) seeds by the method described by Suresh Kumar et al. [7]. Trypsin inhibitor-cellobiose (TIC) and trypsin inhibitor-melibiose (TIM) were prepared by coupling cellobiose or melibiose to soybean trypsin inhibitor through reductive amination [8].  Yeast glycoprotein was isolated from baker’s yeast [9]. Reported procedures were employed to prepare affinity-purified samples of anti-Gal (APAG) [10] and ABG (APABG) [11] from outdated human plasma collected from Department of Transfusion Medicine of this institute in accordance with the Institutional Ethics Committee clearance (IEC/674). Purified samples of AOP1, AOP2, anti-Gal, ABG and human serum albumin (HSA) without non-covalently adhering presence of each other were isolated by electroelution following alkaline electrophoretic separation of APAG or APABG [1]. Anti-Gal, ABG and albumin (HSA) were labeled with FITC as described by Hudson and Hay [12].  Platelets were counted using Horiba ABX Pentra 60 C+ hematology analyzer.