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Anti-Ubiquitin, 64-76 (free/bound) Antibody (Previously Covance catalog# SIG-39400)

Pricing & Availability
Clone
5-25 (See other available formats)
Regulatory Status
RUO
Other Names
Polyubiquitin-C, Polyubiquitin B, RPS 27A, RPS27A, UBA 52, UBA 80, UBA52, UBA80, UBB, UBC, UBCEP 1, UBCEP 2, UBCEP1, UBCEP2, Ubiquitin, ubiquitin B
Previously
Signet Catalog# 9400-02
Signet Catalog# 9400-05
Signet Catalog# 9400-10
Covance Catalog# SIG-39400
Isotype
Mouse IgG1, κ
Ave. Rating
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Product Citations
publications
5-25_Ascites_Ubiquitin_1_031919
Western blot of anti-Ubiquitin, 64-76 (free/bound) antibody (clone 5-25). Lane 1: Molecular weight marker; Lane 2: 30 µg of human brain lysate; Lane 3: 30 µg of mouse brain lysate; Lane 4: 30 µg of rat brain lysate. The blot was incubated with a 1:2000 dilution of the primary antibody overnight at 4°C, followed by incubation with HRP labeled goat anti-mouse IgG (Cat. No. 405306). Enhanced chemiluminescence was used as the detection system.
  • 5-25_Ascites_Ubiquitin_1_031919
    Western blot of anti-Ubiquitin, 64-76 (free/bound) antibody (clone 5-25). Lane 1: Molecular weight marker; Lane 2: 30 µg of human brain lysate; Lane 3: 30 µg of mouse brain lysate; Lane 4: 30 µg of rat brain lysate. The blot was incubated with a 1:2000 dilution of the primary antibody overnight at 4°C, followed by incubation with HRP labeled goat anti-mouse IgG (Cat. No. 405306). Enhanced chemiluminescence was used as the detection system.
  • 5-25_Ascites_Ubiquitin_2_031919
    IHC staining of anti-Ubiquitin, 64-76 (free/bound) antibody (clone 5-25) on formalin-fixed paraffin-embedded Alzheimer's disease human brain tissue. Following antigen retrieval using Sodium Citrate H.I.E.R., the tissue was incubated with a 1:1000 dilution of the primary antibody overnight at 4°C. BioLegend´s Ultra-Streptavidin (USA) HRP kit (Multi-Species, DAB, Cat. No. 929901) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. The image was captured with a 40X objective. Scale bar: 50 µm
  • 5-25_Ascites_Ubiquitin_3_031919
    IHC staining of anti-Ubiquitin, 64-76 (free/bound) antibody (clone 5-25) on formalin-fixed paraffin-embedded Alzheimer's disease human brain tissue. Following antigen retrieval using Sodium Citrate H.I.E.R., the tissue was incubated with a 1:5000 dilution of the primary antibody overnight at 4°C. BioLegend´s Ultra-Streptavidin (USA) HRP kit (Multi-Species, DAB, Cat. No. 929901) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. The image was captured with a 40X objective. Scale bar: 50 µm
  • 5-25_Ascites_Ubiquitin_4_081619
    Western blot of anti-Ubiquitin, 64-76 (free/bound) antibody (clone 5-25). Lane 1: Molecular weight marker; Lane 2: 20 µg of Drosophila head lysate; Lane 3: 20 µg of Drosophila S2 (embryonic) cell lysate. The blot was incubated with a 1:500 dilution of the primary antibody overnight at 4°C, followed by incubation with HRP-labeled goat anti-mouse IgG (Cat. No. 436302). Enhanced chemiluminescence was used as the detection system.
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834901 200 µL 1141 CHF
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Description

Ubiquitin is a small (8.5 kD) regulatory protein that is ubiquitously expressed in tissues of eukaryotic organisms. There are four genes in the human genome that produce ubiquitin; UBB, UBC, UBA52 and RPS27A. UBA52 and RPS27A genes code for a single copy of ubiquitin fused to the ribosomal proteins L40 and S27a, respectively. The UBB and UBC genes code for polyubiquitin precursor proteins.

Ubiquitination is a post-translational modification where a ubiquitin subunit is attached to a protein. Addition of ubiquitin can signal for degradation via the proteasome, alter cellular location, promote or prevent protein interactions, or affect activity. Ubiquitination is carried out stepwise by ubiquitin-activating enzymes (E1s), ubiquitin-conjugating enzymes (E2s), and ubiquitin ligases (E3s), respectively. The cascade results in ubiquitin bound to lysine residues on the protein or to the amino group of the protein's N-terminus.

Ubiquitinations can be either a single ubiquitin unit (monoubiquitination) or a chain of ubiquitin (polyubiquitination). Only poly-ubiquitination on defined lysines, mostly on K48 and K29, is related to degradation with the proteasome, while other polyubiquitinations (e.g. on K63, K11, K6) and monoubiquitinations may regulate processes such as endocytic trafficking, inflammation, translation and DNA repair.

A frameshift mutation in ubiquitin B can result in a truncated peptide missing the C-terminal glycine. This abnormal peptide, known as UBB+1, has been shown to accumulate selectively in Alzheimer's disease and other tauopathies.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Mouse, Rat, Drosophila
Antibody Type
Monoclonal
Host Species
Mouse
Preparation
Ascites
Concentration
The concentration is not quantified as this product is sold as undiluted crude mouse ascites fluid. The concentration might vary from lot-to-lot and an estimated concentration would be 1-3 mg/ml.
Storage & Handling
Store at -20°C or below. Upon initial thawing, apportion into working aliquots and store at -20°C or below. Avoid repeated freeze-thaw cycles to prevent denaturing the antibody. Do not store in frost-free freezers.
Application

WB - Quality tested
IHC-P - Verified
Recommended Usage

Each lot of this antibody is quality control tested by Western blotting. For Western blotting, the suggested use of this reagent is 1:1000 - 1:5000 in human, mouse and rat and 1:500-1:2000 in Drosophila. For immunohistochemistry on formalin-fixed paraffin-embedded tissue, a dilution range of 1:1000 - 1:5000 is suggested. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

This antibody detects ubiquitin and ubiquitinated paired helical filaments (PHF). It is specific for residues 64-76 of ubiquitin and recognizes free and bound ubiquitin, and native or detergent extracted PHF.

Reactivity to Drosophila was only verified with the purified format.

Application References

(PubMed link indicates BioLegend citation)
  1. Grundke-Iqbal I, et al. 1986. J Biol Chem. 261:6084-6089. (WB)
  2. Kudo T, et al. 1994. Brain Res. 1:1-7. (ELISA)
  3. Xu G, et al. 2012. PLoS One 11:e49021. (WB) PubMed
  4. Wang J, et al. 2003. Hum. Mol. Genet. 21:2753. (WB)
  5. Bazzaro M, et al. 2006. Cancer Res. 7:3754. (WB) PubMed
  6. Tabaton M, et al. 1991. Proc. Natl. Acad. Sci. USA 6:2098. (EM, IHC)
  7. Merz PA, et al. 1992. Am. J. Pathol. 2:343. (EM)
Product Citations
  1. Bazzaro M, et al. 2006. Cancer Res. 66:3754-3763. PubMed
  2. Xu G, et al. 2012. PLoS One. 7:e49021. PubMed
RRID
AB_2565315 (BioLegend Cat. No. 834901)

Antigen Details

Structure
Ubiquitin is a 76 amino acid protein with a molecular mass of 8.5 kD.
Distribution

Found in almost all tissues of eukaryotic organisms.

Function
Regulatory protein.
Biology Area
Cell Biology, Neurodegeneration, Neuroscience, Neuroscience Cell Markers, Protein Trafficking and Clearance
Molecular Family
Autophagosome Markers, Protein Modifiers
Gene ID
7316 View all products for this Gene ID
UniProt
View information about Ubiquitin 64-76 on UniProt.org
Go To Top Version: 3    Revision Date: 08.16.2019

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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