If band of conjugate of toxin is exactly behind the standard protein band rather than being ahead in the gel. can we predict conjugation is successful? I have conjugated a toxin to BSA and OVA, both conjugates bands are exactly behind standard protein bands. Please comment about the result.
You mean that the protein band after the conjugation runs slower on the gel than it did before? This would be consistent with an increase in molecular weight due to attachment of the toxin, but could also be due to a change in charge caused by attachment of one or more molecules of the crosslinker. To be really sure of your product you should measure its mass by mass spectrometry.
Dear Professor, this is a new toxin. For others I have got band of conjugate ahead of band of standard protein but for this toxin the band of conjugate is exactly behind (run shorter) than protein.
If the toxin increases the negative charge on the protein, the protein might run faster on the gel (like a smaller protein).
Dear Professor if the conjugate runs shorter (exactly behind standard), can it produce antibodies?
If the toxin is conjugated to the protein and is antigenic then it should be possible to raise antibodies to it (unless it's so toxic that it kills the animals or cells). The main thing is to make sure the toxin is really conjugated to the protein. Mass spec is a good way to do that.
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