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[11] from outdated human plasma collected from Department of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.