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!
Data courtesy of Akaike Lab, Tokyo Institute of Technology
Chemiluminescence detection with Chemi-Lumi One L
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.
If antibody remains on the membrane, it can be detected after the new antigen-antibody reaction causing false result.