Ubiquitin K48 Linkage Antibody

Detection of Human Ubiquitin by Western Blot.

Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line untreated (-) or treated (+) with MG132. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Ubiquitin K48 Linkage Monoclonal Antibody (Catalog # A-101) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Ubiquitin at approximately 75-250 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human Ubiquitin by Western Blot.

25 ng of each linkage of recombinant diubiquitin was run on a 10-20% SDS-PAGE gel prior to blotting on PVDF membrane. Western blots were developed using anti-K48 mAb (A-101, upper panel) or anti-ubiquitin mAb (MAB701, lower panel) primaries at 0.5 µg/ml. The appropriate HRP-labeled anti-rabbit or anti-mouse (R&D Systems HAF008 or HAF007) secondary antibodies were used at a 1:2000 dilution. A single band of appropriate size was detected only in the K48-linked diubiquitin lane using A-101.