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. Author manuscript; available in PMC: 2008 Jun 10.
Published in final edited form as: Immunity. 1996 Jan;4(1):77–85. doi: 10.1016/s1074-7613(00)80300-3

Figure 2. Cross-Linking p58 with Antibody in NK Clones Induces Tyrosine Phosphorylation of a 58 kDa Protein.

Figure 2

Three NK clones expressing the GL183 determinant (SR70, SR47, and SR64) were pooled. The cells (3 × 106 per condition) were preincubated in the absence of primary antibody (minus), with F(ab′)2–GL183 (F–GL183), or intact GL183 (GL183). Following addition of F(ab′)2–goat anti-mouse IgG, cells were incubated for 1 min at 37°C and lysed. GL183 was added to the control samples (minus) after lysis. The GL183 immune complexes (αp58) were collected with mouse anti-goat IgG and the supernatants were subjected to a second immunoprecipitation with anti-phosphotyrosine MAb 4G10 (αptyr). The immunoprecipitated samples were electrophoresed on the same gel and analyzed by Western blot with anti-phosphotyrosine (αptyr). The right panel represents a sequential probing of the membrane in the middle panel using antisera αcyt49. Molecular mass markers are indicated in kilodaltons on the left.