• Validated for Western blot-WB

Anti-ubiquitin antibody (AUB01) was raised against full-length bovine ubiquitin protein. It detects a broad profile of ubiquitinated proteins and free ubiquitin in cell lysates. AUB01 is purified by Protein G affinity chromatography. AUB01 has been conjugated through its amine groups to the carbohydrate groups of horseradish peroxidase (HRP) to make AUB01-HRP. The clone is licensed from Fred Hutchinson Cancer Research Center; the antibody is made in house by Cytoskeleton.

Host/isotype/clone:

• Mouse/IgG1
• Clone: P4D1

Reactivity:

• Broad cross-species reactivity

Amount of material:

• AUB01-HRP: 1 x 100 µl when reconstituted (250-300 µg per tube)
• AUB01-HRP-S: 1 x 25 µl when reconstituted (62.5-75 µg per tube)

Working concentrations:

• WB 1:4000 dilution 0.63-0.75 µg/ml final 400 ml working volume - full size Ab

Formulation

• Supplied as a lyophilized powder
• Buffer composition after reconstitution in 50% glycerol in water: 50 mM PIPES pH7.0, 5% sucrose, 1% dextran, 50% glycerol, 10 mg/ml BSA

Sensitivity:

• Detects ≤3 ng bovine thymus ubiquitin protein

Specificity:

• In 5 µg of Swiss 3T3 cell lysates, a broad range of ubiquitinated proteins and free ubiquitin are detectable. Treatment with 10 µM MG132 for 5 h increases ubiquitinated species by 2–3 fold