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VU-1: The Best Anti-Ubiquitin Antibody Around?

By September 25, 2024 No Comments

Experimental Design:

The purpose of the study is to compare the most common ubiquitin antibodies with LifeSensors UV-1 ubiquitin antibody. VU-1 is a mouse monoclonal antibody developed against human ubiquitin. VU-1 has become the gold standard as it is the best antibody that binds to all types of poly-ubiquitin chains with highest affinity and specificity. To perform apples-to-apples comparison it is essential that the amount of the all the antibodies and the antigens are the same. In addition, it is essential that the amount of antigen, different type of ubiquitin antibodies are diluted in the same buffer at the same concentration at VU1.

Reactivity of antibody with di-ubiquitin, tri-ubiquitin and tetra-ubiquitin is evaluated by running western blots. 30 ng of poly-ubiquitins were used per lane on DSD PAGE. VU-1 was probed against all types of chains and blots at 0.2 ug / ml. All comparative antibodies were diluted to the same concentration in PBS buffer and probed for western blot. The only change is for VU-1 transfer membranes were pretreated with 0.5% glutaraldehyde/PBS pH 7.0 for 20 min. All other conditions were the same for other antibodies. The membranes were then blocked for 1 hour in 5% non-fat milk in TBS-T and incubated in primary antibodies overnight in 5% BSA in TBS-T at 4ºC at 0.2 μg/ml of all the antibodies.

  • VU-1 LifeSensors
  • P4d1  Santa Cruz
  • 1b4 Invitrogen
  • 6C1 Santa Cruz
  • FK1 Biotechne
  • FK2 Sigma
  • Ubi1 Novus Bio

More on the data, and head to head comparison: Here

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