Tomorrow I will be doing SDS-PAGE and then transfer onto nitrocellulose membranes, but I don't know if I'll be able to start the actual Western blot procedure. I was wondering, is there a way for me to store my membrane over the weekend after the transfer? If so, what is the best way? In millipore water? Should I wash w/TBST and then block and keep it in block over the weekend?
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#2
Jordan W
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Posted 24 February 2011 - 12:21 PM
I kept my membrane in the block buffer for like a week once and it still worked beautifully.
#3
mdfenko
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Posted 24 February 2011 - 12:22 PM
i would keep it in the blocking solution, at 4C, over the weekend.
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#4
ecgian
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Posted 24 February 2011 - 12:49 PM
Great, thanks! And it should still work fine after keeping it in block over the weekend?
Thanks for your help!
Thanks for your help!
#5
DarwinDuck
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Posted 15 March 2011 - 11:03 AM
How many times should i use my nitrocellulose membrane and shall i save it in ddH2O water in 4oC between probings?? I've noticed that by doing that my proteins dissapear after 3-4 uses..
#6
mdfenko
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Posted 15 March 2011 - 12:56 PM
DarwinDuck, on 15 March 2011 - 11:03 AM, said:
How many times should i use my nitrocellulose membrane and shall i save it in ddH2O water in 4oC between probings?? I've noticed that by doing that my proteins dissapear after 3-4 uses..
you may want to store in blocking buffer between uses instead of water.
talent does what it can
genius does what it must
i do what i get paid to do
genius does what it must
i do what i get paid to do
#7
DarwinDuck
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Posted 16 March 2011 - 12:41 AM
mdfenko, on 15 March 2011 - 12:56 PM, said:
DarwinDuck, on 15 March 2011 - 11:03 AM, said:
How many times should i use my nitrocellulose membrane and shall i save it in ddH2O water in 4oC between probings?? I've noticed that by doing that my proteins dissapear after 3-4 uses..
you may want to store in blocking buffer between uses instead of water.
I'm not stripping my membrane. I've used to in the past, but i 've realized that it is too stringent for proteins...so i'd rather not..
I suppose you mean blocking buffer with BSA...becasue milk goes bad too soon..about 3 days..I will try it!!
Thanx anyway!!
