Co-immunoprecipitation of CD45 with CD100 in Eskol cells.

Cells lysates were immunoprecipitated with different mAbs: BD16 (anti-CD100), an anti-CD2 (O275) as a negative control, and an anti-CD22 (a molecule known to associate with CD45) and an anti-CD45 (O509) as positive controls. Immunoprecipitates (IP) and cell lysate were subjected to SDS-PAGE, and proteins were transferred on PDVF membrane and blotted with a mixture of anti-CD45 mAbs (O509 and BJ45). Immunoblotted proteins were revealed with goat anti-mouse horseradish peroxidase complex and a system of enhanced chemoluminescence. For detailed experimental procedures, see “Materials and Methods.” We show 2 distinct Western blots: (A) Western blot exposed for 1 minute. (B) Western blot in which cell lysate and CD45 IP were exposed for 5 seconds while CD100 and CD22 IP were exposed for 30 seconds.