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Destain after PoneauS - (Aug/06/2009 )

Hi all,

just a quick question. Is there a way to destain a membrane after I have stained it with Ponceau S after blocking? I have some membranes sitting in PBS in the fridge and I did them quite some time ago when I was learning WB. The problem is that then I stained them with Ponceau S after I had blocked them. Is there a way to get rid of the pinkish color tha obscoures tha result of Ponso?

thank you all

-Aris-

Aris on Aug 6 2009, 03:46 PM said:

Hi all,

just a quick question. Is there a way to destain a membrane after I have stained it with Ponceau S after blocking? I have some membranes sitting in PBS in the fridge and I did them quite some time ago when I was learning WB. The problem is that then I stained them with Ponceau S after I had blocked them. Is there a way to get rid of the pinkish color tha obscoures tha result of Ponso?

thank you all




Rinse a few times in dH2O.

-Dr Teeth-

Rinse a few times in dH2O.



Yes, I have tried that but it always leaves a pinkish color on the blot...

-Aris-

The pink is left on the protein - add your block, it will come out just fine.

-bob1-

I won't come out completely until you strip the blot (I believe it's merely because of intense washing). Ponceau stain supposed not to affect your result, don't worry about pinkish color...

-minylim-

Rinse the membrane with PBS if you really want it to become clear

-bullfrog-

Use 0.1% NaoH in PBS, ponceu will fade like magic. i usually add 10Ál of 1M Naoh to 50ml of PBS containing the membrane and shake it couple of times and its gone. it does not affect post processing of the membranes (i saw it in one of the data sheet from sigma). however you cant destain after the membrane has been exposed to the milk-blocking solution.

-rajgene-