Rabbit F(ab')2 Anti-Rat IgG(H+

E L I S A w i t h p l a t e l e t s

for 1 hour with a 1 :1000 dilution of rat IgG2 a anti-mouse kappa chain monoclonal antibody conjugated to horse-radish peroxidase (LO-MK1 , LO/IMEX) reveal with o-phenylenediamine dihydrochloride (OPD) in revelation solution. SOLUTIONS

Purification of mAb (IgG)

7.8 or Binding Buffer.(6) BD tubing 14-170-12F (Fisher).(7) Polyethylene tubing Clay Adanes.(8) 18 Gauge needles.(9) Binding Buffer (BioRad): make 1L. 314g/L ddH2O and filter through 0.22 um, filter. pH=9.0.(10) Elution buffer (BioRad): make 500 ml. 11g/500 ml

Purification of mAb (IgG)

buffer (pH9.0) ~ 200 ml. Filter protein mixture Add protein mixture or culture supernatant containing Ab (adjust pH to 7.8 with Binding buffer ; red color) to the Protein A column. Mouse antibodies of the IgG1 subclass