ProtoLift Western Stripping Buffer Protocol

Stripping Procedure
 
CAUTION: This procedure requires the use of mercaptoethanol, a hazardous substance. Work in a fume hood.
 
  1. Probe the membrane using your preferred protocol. (do not allow the membrane to dry before stripping).
     
  2. Rinse the membrane in two washes of PBS or TBS. 
     
  3. Prepare a working ProtoLift Western Stripping Buffer solution by adding 2-mercaptoethanol to 0.7% (v/v). To prepare 10 ml of working solution, add 70μl mercaptoethanol to 10 ml ProtoLift stock solution.
     
  4. Place the membrane in a dish and add enough working solution to completely immerse the membrane. Alternatively the membrane can be sealed in a plastic bag with the working solution.
     
  5. Incubate at room temperature for 10 minutes with shaking. PVDF membranes will become transparent in the solution. This is normal and the membrane will return to its regular appearance after the stripping solution is removed.
     
  6. Wash the membrane three times for 10 minutes each with large volumes of PBS or TBS to remove the stripping solution and reducing agent.  The blot is now ready to be reprobed.
 
Note: ProtoLift Western Stripping Buffer is not recommended for nitrocellulose blotting.
Catalog Number: 
EC-889
Full Protocol Sheet: 

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