Primary cells express the TRIM-containing αβTCR. (A) 31.13scTCRβ cells were lysed with digitonin on different days using different lots of digitonin. scTCRs were purified from NP-sepharose affinity columns using free NIP for elution. TCRs were separated by BN-PAGE and immunoblotted against ζ. (B) TCRs were purified from pervanadate-stimulated Jurkat cells as in figure 1A (lane 1). Membranes were prepared from human PBMC and lysed in digitonin (lane 2). Murine thymocytes were stimulated with pervanadate and and TCRs purified with anti-phosphotyrosine antibodies followed by phenylphosphate elution (lane 3). TCRs were separated by BN-PAGE and immunoblotted against ζ. Ferritin was used as a marker protein as in figure 1.