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MABS486 Anti-Ubiquitin (pan) Antibody, clone 2G7B8

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MABS486
100 μg  
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Overview

Replacement Information

Key Spec Table

Species ReactivityKey ApplicationsHostFormatAntibody Type
HWBMPurifiedMonoclonal Antibody
Description
Catalogue NumberMABS486
DescriptionAnti-Ubiquitin (pan) Antibody, clone 2G7B8
Alternate Names
  • Polyubiquitin-B
  • Polyubiquitin-C
  • Ubiquitin
  • Ub
  • Anti-Ubiquitin, UbiSite, clone 2G7B8
Background InformationUbiqutin (Ub) is initially produced as a 229-amino acid Polyubiquitin-B (UniProt P0CG47) precursor protein that is encoded by the UBB gene (Gene ID: 7314) or a 685-amino acid Polyubiquitin-C precursor protein (UniProt P0CG48) encoded by the UBC gene (Gene ID: 7316) in human. Post-translational cleavages yield multiple copies of identical 76-amino acid ubiquitin (also known as high mobility group nonhistone protein HMG-20) molecules. Ubiquitination is one of the most common post-translational modifications (PTMs) of cellular proteins, where Ub is linked covalently via its carboxyl terminus (Gly76) to usually lysine residues in target proteins. In a given target, lysine residue can be linked to one single Ub molecule (monoubiquitylated) or to a chain of Ub molecules (polyubiquitylated). In a polyUb chain, Ub molecules can be linked through one of the seven Lys residues (K6, K11, K27, K29, K33, K48, and K63) or through the Ub N-terminus Met1 residue (which generates linear chains). Lys-C is an endopeptidase that cleaves proteins on the C-terminal side of unmodified lysine residues, but not Ub-modified Lysine residues, generating peptide fragments with a C-terminus Lysine and internal Lysine residue(s) linked to a Ub C-terminal remnant (as a result of Lys-C cleavage between Ub K63 and E64). Antibodies that specifically target this Ub C-terminal remnant (i.e., E64 to Gly76 or E48 to Gly76 if K63 is ubiquitinated in a polyUb chain) are compatible with the patented UbiSite® methodology for Lysine ubiquitination site determination developed by the laboratory of Dr. Blagoy Blagoev at University of Southern Denmark. Such antibodies can isolate Lys-C digested fragments with Ub modification site(s), followed by further trypsin digestion to leave only Gly75-Gly76 covalently linked to the modified Lysine on the isolated peptide fragments prior to Mass spectroscopy (MS) analysis to determine the Lysine modification site(s). They have also use UbiSite® technology to identify serine and threonine residues undergoing HOIL-1 catalyzed ubiquitylation. Threonine residues 12, 14, 22 and serine 20 of ubiquitin are reported to form ester bonds with the C-terminal carboxylate of another ubiquitin molecule thereby increasing the number of ubiquitin linkage types. Serine 175 of IRAK4 and serine 136, 168 and threonine 168 of IRAK2 have been recognized as HOIL-1 catalyzed ubiquitination sites. (Ref.: McCrory, EH., et al. (2022). Biochem. J. 479(23); 2419-2431; Akimov, V., et al. (2018). Nat. Struct. Mol. Biol. 25(7):631-640).
References
Product Information
FormatPurified
PresentationPurified mouse monoclonal IgG1κ antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Quality LevelMQ100
Applications
ApplicationAnti-Ubiquitin (pan)(also known as Anti-Ubiquitin, UbiSite, clone 2G7B8) is a mouse monoclonal antibody that detects Ubiquitin and is used in Western Blotting and Mass spectrometry.
Key Applications
  • Western Blotting
Application NotesWestern Blotting Analysis: 1.0 µg/mL from a representative lot detected the accummulation of ubiquitinated proteins in HEp-2 human epithelial cells following proteasome inhibitor MG-132 (Cat. No. 474790) treatment (Courtesy of Dr. Blagoy Blagoev, University of Southern Denmark).
Biological Information
ImmunogenKLH-conjugated linear peptide corresponding to 13 amino acids from the C-terminal region of human ubiquitin.
EpitopeNear C-terminus.
Clone2G7B8
ConcentrationPlease refer to lot specific datasheet.
HostMouse
SpecificityClone 2G7B8 detects monoubiquitinated (MonoUb) and polyubiquitinated (polyUb) proteins, as well as free ubiquitin (Ub) molecule by targeting Ub sequence C-terminal to Lys63 (UbiSite® tag). Clone 2G7B8 can be used to detect (e.g. by Dot blot) and isolate (i.e. by immunoprecipitation) peptides derived from Ub-modified proteins after endoproteinase Lys-C digestion. Lys-C cleaves Ub and Ub-modified proteins after unmodified Lysine residues, but not Ub-modified Lysine residues, generating peptide fragments with their modified lysine residues linked to Ub C-terminal remnant (UbiSite™ tag) that cannot be recognized by other ubiquitin antibodies whose epitopes lie N-terminally to Lys63. The isolated peptide fragments can then be subjected to Mass spectroscopy (MS) analysis to allow identification of ubiquitin-modification sites.
IsotypeIgG1κ
Species Reactivity
  • Human
Species Reactivity NoteHuman. Broad species reactivity expected based on 100% sequence homology.
Antibody TypeMonoclonal Antibody
Entrez Gene Number
Gene Symbol
  • UBB
  • UBC
Purification MethodProtein G Purified
UniProt Number
Molecular WeightVariable depending on the size of the ubiquitinated protein(s)/peptide(s) and the degrees of ubiquitination.
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality AssuranceEvaluated by Western Blotting in HEp-2 human epithelial cell lysate.

Western Blotting Analysis: 2.0 µg/mL of this antibody detected ubiquitinated proteins in 10 µg of HEp-2 human epithelial cell lysate.
Usage Statement
  • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage and Shipping Information
Storage ConditionsStable for 1 year at 2-8°C from date of receipt.
Packaging Information
Material Size100 μg
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalogue Number GTIN
MABS486 04055977315783

Documentation

Anti-Ubiquitin (pan) Antibody, clone 2G7B8 SDS

Title

Safety Data Sheet (SDS) 

Anti-Ubiquitin (pan) Antibody, clone 2G7B8 Certificates of Analysis

TitleLot Number
Anti-Ubiquitin (pan) Antibody, UbiSite®, Clone 2G7B8 - 4124922 4124922
Anti-Ubiquitin (pan), clone 2G7B8 3074312
Anti-Ubiquitin (pan), clone 2G7B8 - 3172375 3172375
Anti-Ubiquitin (pan), clone 2G7B8 - 3246169 3246169
Anti-Ubiquitin (pan), clone 2G7B8 - 3328863 3328863
Anti-Ubiquitin (pan), clone 2G7B8 - 3540374 3540374
Anti-Ubiquitin (pan), clone 2G7B8 - 3821557 3821557
Anti-Ubiquitin (pan), clone 2G7B8 - 3988914 3988914
Anti-Ubiquitin (pan), clone 2G7B8 -Q2648348 Q2648348
Anti-Ubiquitin (pan), clone 2G7B8 Monoclonal Antibody 3099487