![Figure 3. The BH1 domain mediates the interaction between anti-Bcl-2-scFv-TAT and Bcl-2. (A) Binding of anti-Bcl-2-scFv-TAT to intracellular Bcl-2. Bcl-2 was immunoprecipitated from RBL cells with anti-Bcl-2 immobilized on protein A-agarose. The binding of anti-Bcl-2-scFv-TAT (lane 1) or nonspecific scFv-TAT (lane 2) to intracellular Bcl-2 was determined by Western blot analysis using antibody against the myc tag on the scFv. (B) The sequence of the Bcl-2-BH domains and their homology to other BH domains in the Bcl-2-family proteins. The box encloses the BH domains, and the Bcl-2 sequences before and after the box complete the fragments for PCR. (C) Bcl-2 fragments BH1, BH2, BH3, and BH4 were labeled with [35S]-methionine in vitro and added to anti-Bcl-2-scFv-TAT immobilized on agarose beads. Retained [35S]-labeled fragments were determined by SDS-PAGE and autoradiography. One representative experiment of 3 is shown.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/102/7/10.1182_blood-2002-12-3921/6/h81935002003.jpeg?Expires=1735541997&Signature=DKl6OrnAFCjIy1oOEvztZU-W65YCIlDtuPADlAeGbXIFUI143txgu0aJo2aa6GBqgbP3T6KHJwdznluZDPHjwEQFv0Fa7E~Ms8gzRb1jNAGSu4~bFQoIPlzXQx1MIGB5Ym27lavTbBqWccMMnBVJ4ql6OsNTc9LXJjkoig8x6aExCJxm5b1vL~SXHr4gHxJfgEl7370vrkKfKJNDb2uBjFUtzjqIqMkClapphrrWS-zEW7ZCYvOSucZZci4-GRpUToB6tcwQ8hU2lfKMQ1daTX0DRuTQt2ST4xltzMlIVKWwRrK7Q90eavaE7D8SKCJvBXIpq062nPt~gyp1Pyl~qg__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
The BH1 domain mediates the interaction between anti-Bcl-2-scFv-TAT and Bcl-2. (A) Binding of anti-Bcl-2-scFv-TAT to intracellular Bcl-2. Bcl-2 was immunoprecipitated from RBL cells with anti-Bcl-2 immobilized on protein A-agarose. The binding of anti-Bcl-2-scFv-TAT (lane 1) or nonspecific scFv-TAT (lane 2) to intracellular Bcl-2 was determined by Western blot analysis using antibody against the myc tag on the scFv. (B) The sequence of the Bcl-2-BH domains and their homology to other BH domains in the Bcl-2-family proteins. The box encloses the BH domains, and the Bcl-2 sequences before and after the box complete the fragments for PCR. (C) Bcl-2 fragments BH1, BH2, BH3, and BH4 were labeled with [35S]-methionine in vitro and added to anti-Bcl-2-scFv-TAT immobilized on agarose beads. Retained [35S]-labeled fragments were determined by SDS-PAGE and autoradiography. One representative experiment of 3 is shown.
