Ubiquitin Antibody (1001C) [Unconjugated] - K48 Linkage

Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line untreated (-) or treated (+) with MG132. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Ubiquitin K48 Linkage Monoclonal Antibody (Catalog # A-101) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Ubiquitin at approximately 75-250 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

25 ng of each linkage of recombinant diubiquitin was run on a 10-20% SDS-PAGE gel prior to blotting on PVDF membrane. Western blots were developed using anti-K48 mAb (A-101, upper panel) or anti-ubiquitin mAb (MAB701, lower panel) primaries at 0.5 µg/ml. The appropriate HRP-labeled anti-rabbit or anti-mouse (R&D Systems HAF008 or HAF007) secondary antibodies were used at a 1:2000 dilution. A single band of appropriate size was detected only in the K48-linked diubiquitin lane using A-101.

Product Discontinued

Ubiquitin Antibody (1001C) [Unconjugated] - K48 Linkage Summary

• Additional Information: Recombinant Monoclonal Antibody. Product is in the process of being discontinued.
• Immunogen: K48-linkage from human ubiquitin
  Accession # P0CG47
• Specificity: This antibody detects endogenous, human proteins containing K48-linked polyubiquitin chains in Western blots. This antibody detects purified, recombinant K48-linked polyubiquitin chains, but has no cross-reactivity to monoubiquitin or polyubiquitin of other linkages
• Source: N/A
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Gene: RPS27A
• Purity Statement: Protein A or G purified from cell culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 3 ug/mL
  • Western Blot 0.5 ug/mL

Packaging, Storage & Formulations

• Storage: Store the unopened product at -20 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Storage below -20 °C is not recommended. Do not use past expiration date.
• Buffer: 0.5 mg/mL in PBS, pH 7.4, 50% glycerol, 0.09% sodium azide
• Preservative: 0.09% Sodium Azide

Notes

Not for use in humans

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Ubiquitin Antibody (1001C) [Unconjugated] - K48 Linkage

• RPS27A
• UBA52
• UBB ubiquitin B
• UBB
• UBC
• Ubiquitin

Background

Polyubiquitin chains are composed of ubiquitin monomers that are covalently linked through isopeptide bonds (other than linear, or “Met1-linked” polyubiquitin). Isopeptides are formed between a lysine residue of one Ubiquitin molecule and the C­terminal glycine residue of another Ubiquitin molecule. Seven of the seventy-six amino acids in ubiquitin are lysine residues that can participate in polyubiquitin chain formation. Linkage through specific lysine residues is thought to serve as a signal that affects protein degradation, signaling, trafficking, and other cellular processes. K48-linked polyubiquitin chains attached to substrate proteins often serve as a recognition sequence for targeting and destruction of the substrate by the 26S Proteasome. This antibody detects the K48 linkage. It does not detect monoubiquitin or ubiquitin liked via any other lysine residue. Reactivity across all species is anticipated.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Ubiquitin Antibody (A-101)(2)

Publications using A-101

• J Wang, T Zhang, CJ Johnston, SY Kim, MJ Gaffrey, D Chalupa, G Feng, WJ Qian, MD McGraw, C Ansong Protein thiol oxidation in the rat lung following e-cigarette exposure Redox Biol, 2020-10-10;37(0):101758. 2020-10-10 [PMID: 33080441] (Western Blot, Rat)
• M Jalovecka, D Hartmann, Y Miyamoto, L Eckmann, O Hajdusek, AJ O'Donoghue, D Sojka Validation of Babesia proteasome as a drug target Int J Parasitol Drugs Drug Resist, 2018-08-07;8(3):394-402. 2018-08-07 [PMID: 30103207] (Western Blot)

Bioinformatics

• Gene Symbol: RPS27A
• Uniprot:
  • P0CG47()