I am planning to develop a multispecies (for marsupials) indirect ELISA and looking for a suitable antibody binding reagent. To my knowledge Protein A/G and protein L seem to be good candidates, but protein L appears to have a better binding to many marsupial immunoglobulins than protein A/G based on the immunoblotting results. However, Protein L binding is restricted to antibodies that contain the right subtypes of kappa light chains. Please let me know your experience/suggestions for using protein L in ELISAs.
Tharaka Liyanage Not sure about the structure of marsupial antibodies, is there Fc or a homologue? Antibodies against that region, conjugated to a reporter like HRP should do the trick. Or try Protein A/G/L conjugates with HRP.
Wolfgang Schechinger Yes they have similar antibodies to other mammals, except they dont have IgD. I thought to use protein A/G initially but for some marsupial species it not binding well with their antibodies. I have looked into A/G/L conjugate as well, but I could only find unconjugated protein A/G/L and couldn't find anything that is conjugated with HRP.
Then conjugate yourself. Find "Harlow and Lane - using Antibodies" or their "Antibodies - A Laboratory Manual" in the library, get a HRP with a high RZ value and go ahead.
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