Ubiquitin-HRP Antibody Mouse Monoclonal

Ubiquitin-HRP Mouse Monoclonal Antibody

Mouse / IgG1


Species Reactivity
Wide range of species

Validation Data
Ub Antibody White Paper



Anti-ubiquitin-HRP antibody is a pan-ubiquitin, mouse monoclonal antibody conjugated with HRP 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-HRP has been validated in western blot applications and has been shown to be more potent than unconjugated AUB01 used with standard mouse-HRP secondary.  AUB01 is purified by Protein G affinity chromatography and is supplied as a lyophilized white powder.

Validated Applications

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. 

Ubiquitin Western Blot

Western Blot using Ubiquitin-HRP Antibody

Untreated and MG-132 treated cell lysate were separated by SDS-PAGE and transferred to PVDF. Western blot using an unconjugated (AUB01, 1 µg/ml) or HRP-conjugated (AUB01-HRP, 1 µg/ml) ubiquitin antibody was performed. Anti-Mouse HRP secondary was used for AUB01 but not AUB01-HRP.

Ubiquitin Western Blot

Ubiquitin Background

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.

For more information contact:  signalseeker@cytoskeleton.com

Associated Products:

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-HRP-S):  


Certificate of Analysis:  available upon request

AuthorTitleJournalYearArticle Link
Horita, Henrick et al.Identifying Regulatory Posttranslational Modifications of PD-L1: A Focus on MonoubiquitinatonNeoplasia (United States)2017ISSN 1476-5586

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AuthorTitleJournalYearArticle Link
Horita, Henrick et al.Identifying Regulatory Posttranslational Modifications of PD-L1: A Focus on MonoubiquitinatonNeoplasia (United States)2017ISSN 1476-5586