I am doing an ELISA and my peptide will be conjugated to HSA. I am screening human serum for the antibody of interest. Will the HSA react with the human serum? If so, I need a suitable alternative to HSA that will be affordable and easy to use.
Thanks.
I would not use HSA, depending of purity, it may react with your serum.
BSA, dextrans, etc are better alternatives.
A Colleague of mine says that BSA may also react to the human sera. Does dextran immobilize well on ELISA plates? I saw your paper on the 9 immunodominant proteins where you had coupled to BSA but this was for the lateral flow test. Cool paper! Maybe BSA would be a better choice as a negative control can eliminate background?
Thanks.
Thank you Natasha,
We routinely run ELISAs with BSA conjugates. In the US the incidence of BSA cross-reacting samples is low. In my lab the evidence is anecdotal at best, perhaps one sample in a thousand, but you always find samples that react with pretty much anything. In the past we have found samples that react with PEG for example.
We have found that Dextrans bind well to medium and high binding plates. You can even find modified Dextrans in the market with amines in them that you can use for coupling.
Good luck
Hi Natasha,
HSA and BSA should be good scaffolds the prepare conjugates I agree with Gonzalez that interference and cross-reacting samples are low and you will always find such samples.
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