Published TCIMAIL newest issue No.197
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Antibody Stripping Solution for Chemiluminescent Western Blotting
Western blot stripping buffer (1) is a reagent to remove antibodies from membranes after Western blotting detection using chemiluminescence.1,2) Depending on the affinity of antigen and antibody, the antibody can be removed from the membrane by soaking the membrane in 1 after detection and allowing it to react for 30 minutes at room temperature. Because antigens on the membrane are retained after removal of antibodies, detection can be performed again using another primary antibody. In addition, the advantage is that multiple antigens can be detected in a single electrophoresis and transfer, saving sample and experimental time.
Figure. Two-fold dilution series of HeLa cell lysate were separated by SDS-PAGE (5 - 0.078 µg/lane).
- (A) First detection. Detected by binding anti-αTubulin antibody (Rabbit IgG).
- (B) Antibodies were stripped using stripping buffer from 1or other company product.
- (C) Second detection. The stripped membrane was blocked again and detected by binding with anti-GAPDH antibody (Mouse IgG).
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References
- 1) An overview of technical considerations for Western blotting applications to physiological research
- 2) Shc, Grb2, Sosl, and a 150-Kilodalton Tyrosine-Phosphorylated Protein Form Complexes with Fms in Hematopoietic Cells
