I want to make an indirect ELISA (Ag+anti-Ag+anti-anti-agHRP). And I want to reconstitute the lyophilized antigen (peptides). The question is, what preservative should be used for Ag storage , if it is known that sodium azide blocks the HRP?. I doubt if anyway I can use sodium azide, because washes are performed before adding the Ab-HRP (Wash Buffer contains no sodium azide).
You are right, the washes will remove the sodium azide, so when you treat your wells with anti-anti-agHRP there should be no sodium azide blocking HRP. After you coat your plate with your Ag (+ sodium azide), then wash the plate with the washing buffer, treat the wells with your anti-Ag antibody, then wash again, and then treat the plate with your 2nd Ab-HRP. Two cycles of washing should be enough to remove all sodium azide.
Cygnus' ELISA kits use HRP and they ship with something called Proclin 300 in the antibodies and kit standards. I looked it up and it's available from Sigma. http://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/SAFC/General_Information/proclin_300_features_benefits.pdf
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