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WB Stripping Buffer Solution

Stripping buffer solution

WB Stripping Solution removes conjugated antibodies from blots, enabling subsequent detections with different antigens on the very same blotting membrane. After the first antigenantibody reaction and following chemiluminescent visualization, the antibody can be removed by the WB Stripping Solution. A second antigen-antibody reaction can be conducted on the same blotting membrane. The same blotting membrane can be probed 2-5 times if chemiluminescent detection is employed. Alternately, one blot can be used multiple times under different conditions to optimise results of antigen-antibody reactions!

  • No heating: reaction at room temperature
  • No odor: does not contain 2-mercaptoethanol
  • Fast: stripping time 5-15 minutes
  • Ready to use: one solution in one bottle

Application 1

1. First antigen-antibody reaction
   Blocking        : Blocking One (30 min)  (code No.: 03953-95)
   Wash            :  t-Tris Buffered Saline
   Primary ab      : anti-paxillin (mouse IgG)
   Secondary ab  : anti-mouse IgG-POD
   Detection       : Chemiluminescence Detection Kit (commercially available product)

2. Stripping
   Solution         : WB Stripping Solution
   Condition        : RT, 15 min for conventional protocol

3. Second, different antiogen-antibody reaction
   Blocking         : Blocking One (30 min) (code No.: 03953-95)
   Wash              :  t-Tris Buffered Saline
   Primary ab       : anti-vinculine (mouse IgG)
                       : anti-actin (mouse IgG)
   Secondary ab   : anti-mouse IgG-POD
   Detection        : Chemiluminescence Detection Kit

Data courtesy of Akaike Lab, Tokyo Institute of Technology

Application 2

Chemiluminescence detection with Chemi-Lumi One L

Difference between WB Stripping Solution and WB Stripping Solution Strong

Apply HPR-labeled anti-GST antibody to 5000 ng, 500 ng, 50 ng, or 5 ng (as desired) of c-Myc-GST antigen on a PVDF membrane, then remove the antibody by agitating gently for 10 minutes using one of the following stripping solutions.

After stripping the antibodies and washing the membrane with t-PBS for 2 min, use the chemiluminescence method to detect the HPR-labeled anti-GST antibody remaining on the membrane.

*Image “e” is a result that shows detection of the antigen with HPR-labeled anti-GST antibody on the “d”. The similar result is marked with “a”. Therefore, WB Stripping Solution Strong only stripped antibodies, not antigens.

Proof of Antibody Removal

If antibody remains on the membrane, it can be detected after the new antigen-antibody reaction causing false result.

  • Check whether any labeled antibody complex remains after the stripping by repeated chemiluminescence detection.
  • Check whether any non-labeled (primary) antibody remains by chemiluminescence detection upon repeated reaction with the secondary antibody


  • Depending on the strength of the antigen-antibody reaction, the antibody complex may remain on the membrane after 5-15 minutes stripping treatment at room temperature. It is recommended to optimize the conditions beforehand. If antibodys remain, it is recommended to extend the stripping time (30 minutes to several hours) and/or raise temperature (37 to 60 oC). Perform blocking again if necessary.
  • The product is an acidic solution (pH 2-3). Wear protective chemical-resistant clothes, gloves, and safety eye-wears. If the product accidentally comes into contact with skin, thoroughly wash the skin with water. Contact a physician if necessary.
  • Use the product after it has been equilibrated at room temperature.
  • This product is not suitable for use for blotting membrane stained with TMB, DAB, 4-chloronaphtol or other visually detectable staining, because these stains will not disappear from the membrane.
  • After use, the product shall be neutralized and discarded.
  • If the product is mixed with WB Stripping Solution Strong (Code No.:05677-65), a white precipitation will occur. When both products are used in turns, the blotting membrane must be washed throughly with a proper buffer or deionized water for 3-5 times. Use separate trays for different reagents.


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Ordering Information

Product Cat.No. Storage PKG Size Price(US$)  
WB Stripping Solution 05364-55 Room Temp. 500 ml 98.00 Buy
WB Stripping Solution Strong 05677-65 4C 500 ml 180.00 Buy
WB Stripping Solution Trial Set (WB Stripping Solution + WB Stripping Solution Strong) 05680-21 4C 40 ml each 30.00 Buy