“The VU-1 antibody from LifeSensors is the best anti-ubiquitin antibody that we have found after trying more than ten different anti-ubiquitin antibodies.”- Aldrin Gomes, Ph.D. Associate Professor University of California, Davis
Ubiquitin (Ub), a highly conserved, 8 kDa polypeptide present in all eukaryotic cells, is conjugated to the ε-amino group of lysine residues in the target protein through the sequential action of three enzymes, an E1 Ub activating enzyme, an E2 conjugating enzyme, and an E3 ligase. In addition, the seven lysines within Ub itself can serve as Ub acceptors leading to the formation of polyubiquitin chains. Among these K48 and K63 linkages are well characterized. K48-linked polyubiquitylation targets proteins for proteasomal degradation whereas K63 linkages regulate signaling events, receptor endocytosis and immune responses. Polyubiquitin linkages at other lysines are less prominent and their physiological role remains largely unknown.
VU-1 is a monoclonal antibody that has been shown to recognize free ubiquitin, K48-, K63-, K11- and linear chains on Western blot. Other linkages have not been specifically tested. VU-1 is an excellent antibody for immunostaining applications generating robust ubiquitin detection and low background with an ability to detect subcellular ubiquitin localization.
|Stability||Stable for weeks at +4°C. Sodium azide can be added to an antibody preparation to a
final concentration 0.02% (w/v).
|Storage||For long term storage (6 to 12 months) store at -20°C. Avoid multiple freeze/thaw cycles.|
- Imunnohistochemical staining: 1:200 – 1:500
- Western blots: 1:1000 – 1:10,000 — Please note: To obtain maximum sensitivity, the membrane should be treated with 0.5% glutaraldehyde prior to exposure to the primary antibody
- Immunohistochemistry using chromogenic substrates with a low backgound (see information on the Datasheet);
- Immunofluorescence in tissue sections and cell culture;
- Western blot analysis to detect mono- and polyubiquitinated proteins as well as free ubiquitin