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Membrane Stripping and Reprobing in Western Blot
Materials
- Wet membrane wrapped or in PBS buffer right after (ECL/Supersignal) detection or stored in a refrigerator (2-8oC)
- TBST (TBS with 0.5% Tween-20).
- Re-Blot Plus Strong Solution (10X) (Chemicon International, Cat# 2504)
- Container for membrane (box for 20/200 ul tips is OK.)
Method
- Dilute 10X Re-Blot Plus Strong Solution with TBST to 1X.
- Strip membrane in the diluted solution at room temperature for 30 min to 60 min depending on antibody affinity.
- Wash with TBST.
- Proceed blocking and reprobing as in regular Western blot procedure
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