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([K48-only]Ub)n-ubiquitinylated substrate

BML-UW0640-0025 25 µg 327.00 USD
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Product Details

Source:Protein components produced in E. coli. Synthesized enzymatically in vitro. (K48-only)Ubn-ubiquitinylated substrate protein was produced using K48-only mutant ubiquitin.
UniProt ID:P0CG47 (UBB), P0CG48 (UBC), P62979 (RPS27A), P62987 (UBA52)
Formulation:Liquid. In 50mM TRIS, pH 7.5, containing 150mM NaCl and 1mM DTT.
Purity:≥95% (SDS-PAGE)
Purity Detail:High molecular weight conjugates were separated from free ubiquitin by size exclusion chromatography.
Quality Control:SDS-PAGE: Multiple high molecular weight bands are observed for (K48-only) Ub conjugate reactions (Fig. 1).
Immunoblotting: Detection of high molecular weight bands with the polyubiquitin-chain reactive monoclonal antibody FK2 (Prod. No. BML-PW8810, 1:1000 dilution).
Application Notes:Uses:
1. Deubiquitinylating enzyme substrates (general/linkage specific).
2. Profiling of DUB linkage-type preference or specificity in combination with other single lysine only polyubiquitinylated substrates (e.g. (K11-only)Ubn-ubiquitinylated substrate, Prod. No. BML-UW0620)
3. Investigation of polyubiquitin chain recognition by and interaction with ubiquitin binding proteins.
Shipping:Dry Ice
Short Term Storage:+4°C
Long Term Storage:-80°C
Use/Stability:Stable for at least 6 months after receipt when stored at -80°C.
Handling:Avoid freeze/thaw cycles. After opening, prepare aliquots and store at -80°C.
Scientific Background:Polyubiquitin chains, linked through specific lysine residues, are useful tools for investigating the specificity and reactivity of deubiquitinylating enzymes (DUBs) and the recognition and interaction of polyubiquitin modified proteins with ubiquitin binding domain (UBDs) containing proteins.
Regulatory Status:RUO - Research Use Only
([K48-only]Ub)n-ubiquitinylated substrate SDS-PAGE
Figure 1: Coomassie stained SDS-PAGE gel showing ubiquitinylation of substrate protein (X) with (K48-only) Ub to give (K48-only)Ubn-ubiquitinylated conjugates (Z; Prod. No. BML-UW0640).
([K48-only]Ub)n-ubiquitinylated substrate Western blot
Figure 2: Western blot showing ([K48-only]Ub)n-ubiquitinylated conjugates (BML-UW0640) detected with polyubiquitin-chain reactive monoclonal antibody FK2 (BML-PW8810, 1:1000 dilution)
([K48-only]Ub)n-ubiquitinylated substrate Western blot
Figure 3: Western Blot showing (K48-only)Ubn-ubiquitinylated substrate (Prod. No. BML-UW0640) incubated with (+) or without (-) USP2cd (Prod. No. BML-UW9850), detected with PAb to ubiquitin-protein conjugates."
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([K48-only]Ub)n-ubiquitinylated substrate SDS-PAGE ([K48-only]Ub)n-ubiquitinylated substrate Western blot ([K48-only]Ub)n-ubiquitinylated substrate Western blot

Product Literature References

Oct4 redox sensitivity potentiates reprogramming and differentiation: Z. Shen, et al.; bioRxiv (2023), Abstract;

General Literature References

Mechanism of ubiquitin-chain formation by the human anaphase-promoting complex: L. Jin, et al.; Cell 133, 653 (2008), Abstract;
Proteasome-independent functions of ubiquitin in endocytosis and signaling: D. Mukhopadhyay & H. Riezman; Science 315, 201 (2007), Abstract;
Intracellular protein degradation: from a vague idea thru the lysosome and the ubiquitin-proteasome system and onto human diseases and drug targeting: A. Ciechanover; Exp. Biol. Med. (Maywood) 231, 1197 (2006), Abstract;
Distinct monoubiquitin signals in receptor endocytosis: K. Haglund, et al.; Trends Biochem. Sci. 28, 598 (2003), Abstract;
The BRCA1/BARD1 heterodimer assembles polyubiquitin chains through an unconventional linkage involving lysine residue K6 of ubiquitin: F. Wu-Baer, et al.; J. Biol. Chem. 278, 34743 (2003), Abstract;
The ubiquitin-proteasome proteolytic pathway: destruction for the sake of construction: M.H. Glickman & A. Ciechanover; Physiol. Rev. 82, 373 (2002), Abstract;

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