5 replies to this topic

[vvmm]

Anybody, please? Thank you!

[bob1]

http://www.sciencedi...003269701956040

[vh04x]

I'm confused. Why not just silver stain the gel before transfering to PVDF?

[vvmm]

I have an opportunity to try “Western SERS” with my proteins, and I don’t have pretty much time to do that. And I already have proteins on PVDF membrane.
There is a protocol for NC membrane, but with PVDF everything is confusing… Of course, I could try with NC, but as I said – don’t have much time.

[ascacioc]

vh04x, on 28 August 2012 - 06:35 PM, said:

I'm confused. Why not just silver stain the gel before transfering to PVDF?

The answer lays in what is your final goal: if you want to send one of your bands for analysis to another lab, it is better to do it by transferring it to PVDF. Moreover, some further analysis is not compatible with NC (e.g. Edmann sequencing)

Andreea

[mdfenko]

vh04x, on 28 August 2012 - 06:35 PM, said:

I'm confused. Why not just silver stain the gel before transfering to PVDF?

silver stained protein will not transfer well. formaldehyde in the developer will "fix" the protein in the gel (yes, it can be reversed, with difficulty, but it's much easier to transfer then stain).