How to detect the antigenicity of peptide? - except ELISA (Jun/28/2006 )

hi,

I want to evaluate the antigenicity of several peptides, but the ELISA assay is not satisfying when human serum is used as primary antibody.

so which other methods can be used to detect the antigenicity of peptides? Thank you!

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Surely, if you're trying to see how antigenic a peptide is, you need to inject it into an organism to see what kind of response it generates... Or do I have it all "bass ackwards"?

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sorry im very new to this feild .....if i have cleaved protein and its immunoreactivity to the antibody is much greater than that of the native form (same amount of protein loaded)...it means that it has greater antigenicity to that antibody please clarify this point to me...

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the antigenic epitope is better exposed and more accessible to the antibody.

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mdfenko, thank you a lot! so for example if i have native and cleaved forms of the same protein antigenic epitope of the cleaved form is better exposed to the antibody than that of the native form.... ...if this is the case.....do you have any idea how to remove this factor? normalize? calculate against some standard?
for example maybe i can calculate anteginicity of the both forms and substract it from the intensity of the band???

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if you have cleaved and then purified the fragment, then when you load the same amount of protein you are loading a greater molar concentration of the fragment. this could account for the greater response in the assay. to normalize you would have to add equimolar amounts of peptide.

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你可以首先把多肽和BSA相偶联, 然后再进行ELISA

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"你可以首先把多肽和BSA相偶联, 然后再进行ELISA"

What do you mean? do I need to incubate the peptide and BSA at 4 degree? how long will be appreciate?

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