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my membranes are blue!


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#1 almost a doctor

almost a doctor

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Posted 19 August 2009 - 12:26 AM

So here's my problem:

Apparently the dish I used to stain my membranes with Ponceu after transfer from SDS-PAGE wasn't as clean as I thought, and had obviously been used for coomassie stain previously, and now my membranes are blue all over!!! :o ;)
I'm optimising a new protocol, so I've used PVDF and nitrocellulose, and both are equally blue, specially round the edges. Thing is I want to scan the membranes once they are stained with Ponceau, and although I did that yesterday the pics are horrible as there's too much blue :(

Any suggestions on how to destain them will be more than appreciated, as water just doesn't seem to do the trick. Can I use methanol and acetic acid as I will do for a gel??

Thanks in advance for any comment. :)

Edited by almost a doctor, 19 August 2009 - 12:32 AM.


#2 lab rat

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Posted 19 August 2009 - 11:40 AM

Hi Doctor,

I wouldn't recommend using methanol and acetic acid to destain the membrane. Just rinse really well with Towbin's. Destaining will take better than an hour, but it won't be complete. You might be able to eliminate enough to stain with Ponceau.

lab rat
42..."An immutable fixed-precision number of unlimited magnitude." <a href="http://en.wikipedia....amming_language)" target="_blank">http://en.wikipedia....amming_language)</a>, accessed 25June2009.




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