MABS486

Anti-Ubiquitin (pan) Antibody, UbiSite^™ ,clone 2G7B8

clone 2G7B8, from mouse

• 別名: Polyubiquitin-B, Polyubiquitin-C, Ubiquitin, Ub, Anti-Ubiquitin,UbiSite^™, clone 2G7B8
• UNSPSCコード: 12352203
• eCl@ss: 32160702
• NACRES: NA.41

特性

• 由来生物: mouse
• 品質水準: 100
• 抗体製品の状態: purified antibody
• 抗体製品タイプ: primary antibodies
• クローン: 2G7B8, monoclonal
• 交差性: human
• テクニック: western blot: suitable
• アイソタイプ: IgG1κ
• NCBIアクセッション番号: NP_001268645.1; NP_066289.3
• UniProtアクセッション番号: P0CG47; P0CG48
• 輸送温度: wet ice
• ターゲットの翻訳後修飾: unmodified
• 遺伝子情報: human ... UBB(7314)

詳細

詳細

Ubiqutin (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).

特異性

Broad species reactivity expected based on 100% sequence homology.

Clone 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.

免疫原

Epitope: Near C-terminus.

KLH-conjugatedlinear peptide corresponding to 13 amino acids from the C-terminal region ofhuman ubiquitin.

アプリケーション

Research Category
細胞シグナル伝達

Research Sub Category
ユビキチン及びユビキチン代謝

Anti-Ubiquitin(pan)(also known as Anti-Ubiquitin, UbiSite^™, clone 2G7B8) is a mouse monoclonalantibody that detects Ubiquitin and is used in Western Blotting and Massspectrometry.

Western 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).

品質

Evaluated 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.

ターゲットの説明

Variable depending on the size of the ubiquitinated protein(s)/peptide(s) and the degrees of ubiquitination.

物理的形状

Protein G Purified

Format: Purified

Purified mouse monoclonal IgG1κ antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

保管および安定性

Stable for 1 year at 2-8°C from date of receipt.

その他情報

Concentration: Please refer to lot specific datasheet.

法的情報

UbiSite is a trademark of Syddansk Universitet (University of Southern Denmark)

免責事項

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.

安全性情報

• 保管分類コード: 12 - Non Combustible Liquids
• WGK: WGK 1
• 引火点(°F): Not applicable
• 引火点(℃): Not applicable