Donkey anti rabbit na934v

Standard SDS-PAGE gel analysis was carried out in a range of gel concentrations. Proteins were either stained with GelCode Blue (ThermoFisher) or transferred to PVDF Immobilon-P transfer membranes (0.45 µM pore size) (Sigma-Aldrich) for immunoblot analysis.^{35 (link)} Antibodies utilized for immunoblotting were: mouse anti-tetraHis (Qiagen, 1:4,000); mouse anti-FLAG M2 (Sigma, 1:10,000); rabbit anti-ubiquitin (Dako, 1:1000); mouse 16B12 anti-HA (Covance, 1:1000); and mouse anti-PGK (yeast phosphoglycerate kinase) (Molecular Probes, 1:20,000). Secondary antibodies used were: sheep anti-mouse NA931V (GE Healthcare, 1:10,000) and donkey anti-rabbit NA934V (GE Healthcare, 1:5,000). Membranes used for anti-His blotting required blocking of nonspecific binding with 3% (w/v) BSA and extensive washing. Other immunoblot analyses used 5% milk for blocking.