I’m trying to measure the level of IgA in breast-milk by ELISA but I’m having problems with the standard (Sandoglobulin) which doesn’t show any colour development after adding the p-Nitrophenyl phosphate (PNPP) substrate but the sample does.
I think Sandoglobulin contains mostly IgG and maybe it can be the cause for not responsing for the ELISA kit.
and always be careful about all the conditions of the test,maybe you made an error during the test.
I also think that sandoglobulin is not the best standard for this elisa, since it mainly contains IgG and traces of IgA and IgM. Try with an IgA standard.
You can use as standard: "IgA from human colostrum" by Sigma-Aldrich as described in the article in attachment; alternatively you can use elisa kit for IgA detection by Binding Site.
Thank you very much for all your comments. I'll go in for the IgA Elisa kit to make my work simple.
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