Substitution of carbonate by acetate buffer for IgG coating in sandwich ELISA.
The first step of sandwich ELISA, namely adsorption of antibodies to plastic microtiter plates, was studied as a function of the pH of the coating buffer. Coating efficiency was assessed in terms of maximum signal (absorbance) observed in ELISA and also estimated by measuring the amount of functional antibodies adsorbed to the plate. While goat antibodies displayed better results after coating with acetate pH 5 buffer, rabbit IgGs generally worked well at pH 7.4. On average, the classical carbonate pH 9.6 buffer was only 50% as efficient.
Cuvelier
A
A
INSERM Unité 295, Faculté de Médecine-Pharmacie de Rouen, France.
Bourguignon
J
J
Muir
J F
JF
Martin
J P
JP
Sesboüé
R
R
D000085:Acetates; D000510:Alpha-Globulins / Q000276:immunology; D000818:Animals; D001666:Binding Sites, Antibody; D002021:Buffers; D002254:Carbonates; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006041:Goats; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000737:chemistry*; D011817:Rabbits; D014361:Trypsin Inhibitors / Q000276:immunology; D000515:alpha 1-Antitrypsin / Q000276:immunology
D000085:Acetates; D000510:Alpha-Globulins; D002021:Buffers; D002254:Carbonates; D007074:Immunoglobulin G; D014361:Trypsin Inhibitors; D000515:alpha 1-Antitrypsin; C011877:inter-alpha-inhibitor