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[Bioscientist]

Hi All

I did a far western blot. I heated my prey proteins with SDS-loading buffer and DTT in one case. I also just heated them in only in the SDS-loading buffer without any DTT. I then ran them on a gel, transferred and used my purified protein as the bait protein.  I got strong bands in the samples that were treated with DTT but not in the samples without DTT.

So what does that mean?

Is this an actual interaction?

Also the place where I got the band, had strong protein bands as I saw in the SDS-PAGE gel. So I was wondering is it possible for bait proteins to bind to some random proteins on the membrane just because it is present in high concentration than the other proteins?

Thanks.

Bioscientist

[mdfenko]

the band strength with the dtt sample buffer is probably due to improved exposure of the epitope to which the antibody was made.