This purified polyclonal antibody reacts with ubiquitin by ELISA and western blot. Although not tested, this antibody is likely functional in immunohistochemistry and immunoprecipitation. For detection of free Ub by western blotting use Tris-Tricine SDS-
Anti-Ubiquitin antibody, generated by immunization with Ubiquitin coupled to Rabbit IgG, was tested by western blot against total cell extract from yeast. Dilution of the antibody between 1:200 and 1:1,000 showed strong reactivity with Ubiquitinated proteins. In this blot the antibody was used at a 1:500 dilution incubated overnight at 4 C in 5% non-fat dry milk in TTBS. Detection occurred using a 1:2000 dilution of HRP-labeled Donkey anti-Rabbit IgG (code 611-703-127) for 1 hour at room temperature. A chemi-luminescence system was used for signal detection (Roche). Other detection systems will yield similar results.
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