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Ubiquitin K48 Linkage Antibody

Catalog #: A-101
2 Citations Datasheet / COA / SDS
Recombinant Monoclonal Antibody. Product is in the process of being discontinued

Discontinued Product

A-101 has been discontinued.
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Detection of Human Ubiquitin by Western Blot.
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Product Details
Citations (2)
FAQs
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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Ubiquitin K48 Linkage Antibody Summary

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
Recombinant Monoclonal Rabbit IgG Clone # 1001C
Purification
Protein A or G purified from cell culture supernatant
Immunogen
K48-linkage from human ubiquitin
Accession # P0CG47
Formulation
0.5 mg/mL in PBS, pH 7.4, 50% glycerol, 0.09% sodium azide
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.5 µg/mL
See below
Immunocytochemistry
Recommended concentration for ICC applications is 3 μg/ml.
 

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Human Ubiquitin antibody by Western Blot. View Larger

Detection of Human Ubiquitin by Western Blot. 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.

Western Blot Detection of Human Ubiquitin antibody by Western Blot. View Larger

Detection of Human Ubiquitin by Western Blot. 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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.

Background: Ubiquitin

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.

Entrez Gene IDs
7314 (Human); 298693 (Rat)
Alternate Names
RPS27A; UBA52; UBB ubiquitin B; UBB; UBC; Ubiquitin

Product Datasheets

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COA
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Product Specific Notices

Not for use in humans

Citations for Ubiquitin K48 Linkage Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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  1. Protein thiol oxidation in the rat lung following e-cigarette exposure
    Authors: J Wang, T Zhang, CJ Johnston, SY Kim, MJ Gaffrey, D Chalupa, G Feng, WJ Qian, MD McGraw, C Ansong
    Redox Biol, 2020-10-10;37(0):101758.
    Species: Rat
    Sample Types: Cell Culture Supernates
    Applications: Western Blot
  2. Validation of Babesia proteasome as a drug target
    Authors: M Jalovecka, D Hartmann, Y Miyamoto, L Eckmann, O Hajdusek, AJ O'Donoghue, D Sojka
    Int J Parasitol Drugs Drug Resist, 2018-08-07;8(3):394-402.
    Applications: Western Blot

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