Technical question about western blot - (Dec/13/2006 )

goodmorning all
i have a quick question how long do u keep ur blocking buffer?
i am using block ace and i reuse it after each block and i do western blotting sometimes evey day and sometimes every 2 days?
is it ok to keep it for one month?
thankx

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Hi, I use each time fresh skim milk poweder (5%) in TBS-T(0,2%). I don't reuse it, I only dilute the antibodies in it, but use everytime fresh one. But mostly because its cheap

Bye

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I use 5% milk powder in TBS1x and store it in the fridge. I only reuse it when it´s 2-3 consecutive days. If I keep it too long even in the frigde, it becames some kind of a yogurt / cheese mixture with an unpleasant smell...

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I keep it only 2 days in the fridge.
I only prepare 20 mL per blot extemporously.

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i prepare each time 5%skimmed milk TBST for blocking.
my colleague who use company blocking buffer does not keep it even for a same-day 2nd blot...

i asume you can keep it for 2-3 consecutive days

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if the blocking buffer contains primary antibody or secondary antibody, i will reuse it 5 times and store it at -20 oc, but pure blocking buffer is use only once.

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we use 5% milk and usually store it in 4C. But before using it always smell it to know if its gone bad or not.

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Hello,

here is a recepie for stable, yoghurt-free blocking buffer...store at -20 degrees and u can reuse it for atleast one month, thaw just before use..

5% BSA in 1X TBS (with 0.2% tween) + 0.002% sodium azide (NaN3)

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thankx everybody
but my blocking buffer is not turning yoghurt or anything in 4 c
i am using Block Ace with tween and sodium azide
and no bacterial contamination also and sometimes i use it for a month or 2
i was just asking from the efficiency point of view.
thankx

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