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Other monoclonal antibodies used in combination with the rabbit anti-myc antibody were: anti-lamin A + B and anti-ubiquitin (Zymed Laboratories, South San Francisco, CA), anti-Hsp70 and anti-Hsp90 (Stressgen, Inc., Victoria, BC, Canada), anti-GFP (Clontech, Inc.
Primary antibodies for actin (clone C4, Boehringer Mannheim Corp., Indianapolis, IN) and ERα (clone h-151, StressGen Biotechnologies Corp, Victoria, BC; 0.1 μg/ml) were also used for western blot analysis as described [ 15 ] .
Negative controls were carried out without primary antibodies and with primary antibodies preabsorbed (100-fold molar excess, 2 h, 25 C) with an 8-oxoguanine oligonucleotide (3850-100-01; Trevigen), human recombinant p53 (Santa Cruz Biotechnology, Santa Cruz, CA), Bcl-2 peptide (amino acids 41-54; StressGen), or recombinant polymerase β (Trevigen).
Purified antibodies to p53 (mouse monoclonal KAM-CC002) and Bcl-2 (rabbit polyclonal AAP-070) were obtained from StressGen Biotechnologies (Victoria BC, Canada).
For dual localization experiments, a 1:1000 dilution of a rabbit polyclonal anti-β-coatomer protein (β-COP) antibody (Affinity BioReagents) or a 1:200 dilution of a rabbit polyclonal anti-protein disulfide isomerase (PDI) antibody (StressGen Biotechnologies) was included with the EE mouse monoclonal.