Figure 2.

Specific binding of Hsp90 in cell extracts to GA affinity beads. Extracts of ³⁵S-labeled SW620 cells (see Fig. 1) were incubated with GA coupled to Affi-Gel 10 (13) or Affi-Gel 10 alone, as indicated, followed by 10 washes in lysis buffer (14) and by elution with SDS buffer. Eluates were analyzed by SDS/PAGE and autoradiography (³⁵S) or immunoblotting with anti-Hsp90 antibody (anti-Hsp90). Total protein (25 μg) was analyzed in lane T. In lanes 1–4, 875 μg of total protein was incubated with 10-μl beads for 60 min at 25°C.