Western blot stripping protocol
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Western blot stripping protocol
Western blot stripping protocol:
Buffer needed:
stripping buffer:
2% SDS
100 mM beta-mercaptoethanol
50 mM Tris, pH 6.8
Procedure:
1. Heat stripping buffer to 50c in water bath.
2. Incubate blot with stripping buffer at 50c for 15-30 min with gentle shaking.
3. Rinse blot several times in TBS.
4. Blot is now ready for re-block and blot.
Last update 29-Aug-2006, Rating Very Good of 5 votes.
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Should I use Tris-base or Tris-HCL to prepare the stripping buffer?
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nguyen: it's 2% by weight, so for example 2 grams of SDS in 100 mL of solution.
100% would be 100 g in 100 mL
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can you tell me how to make the 2%SDS recipe? thank so much!
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Depending on how many times you strip and reprobe, you can still get specific band after 5-7 times stripping of a western blot membrane. PVDF membrane works much better for stripping and reprobing in western blotting.
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How dose it work ? After stripping and reblock, can I still get specific band?
Thanks
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