Hello.
Does anyone know if it's OK to store a washed membrane, after incubation of primary antibody, in TBST at 4oC for about 2 days? I don't have any choice because, I just found out that we're out of the secondary antibody. Thanks.
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#2
merwyn
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Posted 13 June 2009 - 02:45 AM
Hai
fine; try that out and hang the answer in this forum,
fine; try that out and hang the answer in this forum,
#3
Gerard
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Posted 14 June 2009 - 01:09 AM
It will be OK on PBST as long as there is no or very little bacterial or fungal growth.
You can dry the the membrane at the air and store it between filterpaper, my expirience is that even after weeks it works fine after hydratation in PBSt for 15 minutes. This functions well with the protein allone as well with the antibody but I've never tryed with labeling antibodies
You can dry the the membrane at the air and store it between filterpaper, my expirience is that even after weeks it works fine after hydratation in PBSt for 15 minutes. This functions well with the protein allone as well with the antibody but I've never tryed with labeling antibodies
Ockham's razor
Pluralitas non est ponenda sine necessitate
-- "You must assume no plural without necessity".
Pluralitas non est ponenda sine necessitate
-- "You must assume no plural without necessity".
#4
Trewble
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Posted 15 June 2009 - 07:18 AM
Hi,
It's OK to store your membrane in TBST at room temperature or between papers, only if the desired immunodecoration is against protein subunits. If you are dealing with the native protein, i suggest you to keep your membrane at 4ºC in TBST. Nevertheless, it would be advantageous if you incubate again for 1hour with your primary antibody...
Good Luck
Pedro S
It's OK to store your membrane in TBST at room temperature or between papers, only if the desired immunodecoration is against protein subunits. If you are dealing with the native protein, i suggest you to keep your membrane at 4ºC in TBST. Nevertheless, it would be advantageous if you incubate again for 1hour with your primary antibody...
Good Luck
Pedro S
#5
mikew
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Posted 15 June 2009 - 10:46 AM
It should be fine.
I would probably reincubate for 1 hr with the primary.
Could you ask another lab for the secondary?
I would probably reincubate for 1 hr with the primary.
Could you ask another lab for the secondary?
#6
medchemgirl
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Posted 15 June 2009 - 12:19 PM
I agree about the incubating again with ur primary. I've done what u are trying to do and I found out that you can loose in solution some of ur antibodies.
Trewble, on Jun 15 2009, 09:18 AM, said:
Hi,
It's OK to store your membrane in TBST at room temperature or between papers, only if the desired immunodecoration is against protein subunits. If you are dealing with the native protein, i suggest you to keep your membrane at 4ºC in TBST. Nevertheless, it would be advantageous if you incubate again for 1hour with your primary antibody...
Good Luck
Pedro S
It's OK to store your membrane in TBST at room temperature or between papers, only if the desired immunodecoration is against protein subunits. If you are dealing with the native protein, i suggest you to keep your membrane at 4ºC in TBST. Nevertheless, it would be advantageous if you incubate again for 1hour with your primary antibody...
Good Luck
Pedro S
