Technique / Proteomics and protein biochemistry / Western blot assay

Western blot stripping protocol


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 Good of 6 votes.

Write your comment

By kashani on 05-Oct-2008
there is no signal with ECL after stripping but I have good signal with Dab. please help me.
Rating: n/a

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By Guoying Wang on 25-Apr-2008
Should I use Tris-base or Tris-HCL to prepare the stripping buffer?
Rating: Very Good

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By anon on 30-Jan-2008
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

Rating: Good

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By Nguyen on 09-Oct-2007
can you tell me how to make the 2%SDS recipe? thank so much!
Rating: Very Good

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By Joe on 26-Jan-2007
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.
Rating: Very Good

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By gd on 29-Aug-2006
How dose it work ? After stripping and reblock, can I still get specific band?
Thanks
Rating: Good

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