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pH of RIPA buffer??


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#1 yobou

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Posted 21 February 2012 - 01:06 AM

Dear All
I have prepared RIPA buffer with the following ingredients:

50mM TRIS, pH8
150mM NaCl
1% NP-40
0.5% sod. deoxycholate
0.1% SDS

Immediately after buffer preparation I measured its pH and it was 8.35.
I would like to know should I re-adjust the pH back to 8?
one more thing, regarding storage, some members recommend that it should be at -20, but others say that as long as there is no protease or phosphatase inhibitor cocktails it can be stored at refregirator at 4, which is the best temperature for storage?
thanks

#2 GNANA

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Posted 21 February 2012 - 03:12 AM

To be safer i think you can adjust the pH to 8. regarding the storage i always store at 4 degrees that is RIPA without protease and phosphatase inhibitors. i would recommend to add the inhibitors freshly during lysis. hope this helps...

Gnana...
I would prefer being perfectionist rather than a passionist in Research.

I always had an alternate hypothesis....

#3 mdfenko

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Posted 21 February 2012 - 12:16 PM

if you are determining the pH after adding sds then i wouldn't worry about it.

sds makes pH readings questionable (it precipitates in and blocks the porous plug on the electrode due to kcl).
talent does what it can
genius does what it must
i do what i get paid to do




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