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Protocol
Chromophore/Substrate solution (ELISA)
Acc.no:
MXY70G
|
Published:
2008-12-10
by
Unknown user
Keywords: ELISA, Substrate solution, chromophore, OPD
Phosphate-cytrate buffer (pH 5).
Required Materials
- o-Phenylenediamine (OPD) , 0.5 mg/ml
- Cytric Acid , 0.1 M, 98.6 ml (10.357 g/1000 ml)
- Disodium phosphate , 0.2M, 101.4 ml (14.399 g/1000 ml)
- Hydrogen peroxide , 0.02%
Instructions
-
Preparation
Dissolve tablets of OPD (SIGMA) in the appropriate volume of buffer (e.g. one tablet of 60 mg OPD in 120 ml of phosphate-cytrate buffer, pH 5). This solution can be stored at –20°C in appropriate aliquots (6, 12, 18 or 24 ml aliquots, suited respectively for 1, 2, 3 or 4 plates) in the dark. Immediately before use add H2O2 at 0.02% final concentration (e.g. H2O2 30% 4 ?l H2O2 per 6 ml OPD).
Potential carcinogenic agent R45.
Store at 4 °C.
