A 149 residue region (aa 553-702) at the C-terminus of HSP-90 ("HSP-90 antigen") was expressed in E. coli as a GST fusion protein and sent to Noble Life Sciences for anti- body production in rats. After ∼3 mo, we received the antisera, and affinity purified anti-HSP-90 antibodies by use of an MBP- HSP-90 antigen coupled to Affigel matrix (BioRad), using a procedure previously published (Benian et al., 1993). Antibodies work well in western blots, detecting a polypeptide of expected size for HSP-90 (80 kDa), and no detectable extraneous bands at a 1:5000 or 1:2500 dilution.
unc-95