Anti-ubiquitin antibody is a pan-ubiquitin, mouse monoclonal antibody that is part of the Signal-Seeker™ product line. The antibody was raised against full length bovine ubiquitin. The antibody has been shown to recognize poly-ubiquitin, mono-ubiquitin, and free ubiquitin (Fig 1). Ubiquitin is a highly conserved protein and AUB01 is predicted to recognize ubiquitin from a wide range of species (5). AUB01 is purified by Protein G affinity chromatography and is supplied as a lyophilized white powder.
Western Blot using Ubiquitin Antibody
Anti-Ubiquitin Antibody (Cat. # AUB01) was used at a 1:500 dilution. Bovine thymus ubiquitin was run as follows; Lane 1-50 ng, Lane 2-25 ng. Lane 3– 12.5 ng, Lane 4-6.25 ng, Lane 5-3.12 ng. Lanes 6 & 7 represent 20 µg of Swiss 3T3 cell lysate from cells treated for 5h with 10 µM MG132 (Lane 6) or untreated cells (Lane 7). Arrow indicates free ubiquitin band (8 kD), higher molecular weight bands are ubiquitinated proteins. To see the full Western blot protocol, see the product datasheet.
Ubiquitin (Ub) and ubiquitin-like proteins (Ubls, e.g. SUMO, Nedd) are a group of approximately 15 proteins that have a molecular weight of around 8 kD. During the ubiquitination process, these are conjugated via activating (E1), conjugating (E2) and ligating (E3) enzymes to lysines of a target protein (1). Mammalian cells express over 600 potential ubiquitin ligases which exceeds that of the kinase superfamily of PTM proteins (2).
One function of ubiquitination is to target proteins for proteosomal degradation. This role can range from a general housekeeping function that clears miss folded proteins from a cell to involvement in tightly regulated spatio-temporal cell signaling events (1). An emerging function of ubiquitination is its ability to activate proteins via the creation of unique protein:protein interactions (3). In common with many other PTMs, ubiquitination is reversible. Ubiquitin-specific proteases (USPs or DUBs) remove ubiquitins from target proteins (4). The reversible nature of ubiquitination further enhances the potential of this PTM to dynamically regulate protein function.
1) Grabbe, C. et al. 2011. The spatial and temporal organization of ubiquitin networks. Nat. Rev. Mol. Cell. Biol. 12:295-307.
2) Deshaies R.J. & Joazeiro, C.A. 2009. RING domain E3 ligases. Ann. Rev. Biochem. 78: 399-434.
3) Lomeli, H. & Vazquez. 2011. Emerging roles of the SUMO pathway. Cell Mol. Life Sci. 68:4045-4064.
4) Faesen, A.C. et al. 2012. The role of UBL domains in ubiquitin specific proteases. Biochem. Soc. Trans. 40:539-545.
5) Zuin, A. et al. 2014. Ubiquitin signaling: Extreme conservation as a source of diversity. Cells 3:690-701.
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Signal-Seeker™ Ubiquitination Detection Kit (Cat. # BK161)
Signal-Seeker™ Ubiquitin Affinity Beads (Cat.# UBA01-beads)
Signal-Seeker™: BlastR™ Rapid Lysate Prep Kit (Cat. # BLR01)
For product Datasheets and MSDSs please click on the PDF links below.
Sample Size Datasheet (Cat. AUB01-S):
Bulk Size Datasheet (Cat. AUB01-XL):
Certificate of Analysis: Lot 001
For the most recent publications citing this and other Signal-Seeker™ products, see our Signal-Seeker™ Validation Data Page click here
|Wang, Longlong et al.||Disrupting the HDAC6-ubiquitin interaction impairs infection by influenza and Zika virus and cellular stress pathways||Cell Reports||2022|
|Martin, Thomas G. et al.||Cardiomyocyte contractile impairment in heart failure results from reduced BAG3-mediated sarcomeric protein turnover||Nature Communications||2021||ISSN 2041-1723|
|Manzione, Maria Giulia et al.||Co-regulation of the antagonistic RepoMan:Aurora-B pair in proliferating cells||Molecular Biology of the Cell||2020||ISSN 1939-4586|
|Rivas, José et al.||KCTD5, a novel TRPM4-regulatory protein required for cell migration as a new predictor for breast cancer prognosis||FASEB Journal||2020||ISSN 1530-6860|
|Wenzel, H. Jürgen et al.||Astroglial-targeted expression of the fragile X CGG repeat premutation in mice yields RAN translation, motor deficits and possible evidence for cell-to-cell propagation of FXTAS pathology||Acta neuropathologica communications||2019||ISSN 2051-5960|
|Álvarez, Alhejandra et al.||KCTD5 and ubiquitin proteasome signaling are required for Helicobacter pylori adherence||Frontiers in Cellular and Infection Microbiology||2017||ISSN 2235-2988|
|Braganza, Andrea et al.||UBE3B is a calmodulin-regulated, mitochondrion-associated E3 ubiquitin ligase||Journal of Biological Chemistry||2017||ISSN 1083-351X|
|Horita, Henrick et al.||Identifying Regulatory Posttranslational Modifications of PD-L1: A Focus on Monoubiquitinaton||Neoplasia (United States)||2017||ISSN 1476-5586|
|Hukema, Renate K. et al.||Reversibility of neuropathology and motor deficits in an inducible mouse model for FXTAS||Human Molecular Genetics||2015||ISSN 1460-2083|
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