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Di-ubiquitin (K63-linked)

BML-UW0730-0050 50 µg 310.00 USD
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Polyubiquitin chains, linked through specific lysine residues, are useful tools for investigating, amongst other things, the specificity and reactivity of deubiquitinylating enzymes (DUBs) and the recognition and interaction of polyubiquitin modified proteins with by ubiquitin binding domain (UBDs) containing proteins.

Product Details

Alternative Name:Ub2
Source:Protein components produced in E. coli. Synthesised enzymatically in vitro.
UniProt ID:P0CG47 (UBB), P0CG48 (UBC)
Formulation:Liquid. In 20mM TRIS-HCl, pH 7.5, containing 0.15M NaCl, 1mM EDTA.
Purity:≥95% (SDS-PAGE)
Application Notes:For use in deubiquitinylating enzyme assays and polyubiquitin binding studies.

Suggested uses:
  1. Deubiquitinylating enzyme substrates (general/linkage specific).
  2. Investigation of polyubiquitin chain recognition by and interaction with ubiquitin binding proteins.
Shipping:Blue Ice
Short Term Storage:-20°C
Long Term Storage:-20°C
Use/Stability:Stable for at least 12 months after receipt when stored at -20°C. It is recommended that the product be kept at a moderately high protein concentration (>1mg/mL) and in the presence of buffer. If a more dilute reagent is required, the inclusion of an inert carrier protein (e.g. ovalbumin at 0.5-1.0mg/mL) in the storage buffer is recommended in order to minimise loss of the reagent due to absorption.
Handling:Avoid freeze/thaw cycles. After opening, prepare aliquots and store at -20°C.
Technical Info/Product Notes:K63-linked di-ubiquitin (Ub2) is made by incubating E1 and Ubc13-Mms2 with equal concentrations of ubiquitin capped at the future proximal chain terminus (Asp77-ubiquitin) and ubiquitin capped at the future distal chain terminus (K48R/K63R-ubiquitin). The distal terminus of the resulting dimer is de-blocked by treatment with yeast ubiquitin hydrolase-1 (YUH-1) followed by purification by anion exchange chromatography.
Regulatory Status:RUO - Research Use Only
Di-ubiquitin (K63-linked) SDS-PAGE
SDS-PAGE of di-ubiquitin Ub2, K63-linked (Lane 1: Prod. No. BML-UW0730, 5µg), in comparison with Ub3, K63-linked (Lane 2: Prod. No. BML-UW0745, 5µg) and Ub4, K63-linked (Lane 3: Prod. No. BML-UW0715, 5µg)."
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Di-ubiquitin (K63-linked) SDS-PAGE

Product Literature References

Silybins are stereospecific regulators of the 20S proteasome: M. Persico, et al.; Bioorg. Med. Chem. 66, 116813 (2022), Abstract;
The specificity of ubiquitin binding to ubiquilin-1 is regulated by sequences besides its UBA domain: C.A. Harman & M.J. Monteiro; Biochim. Biophys. Acta Gen. Subj. 1863, 1568 (2019), Abstract;

General Literature References

In vitro assembly and recognition of Lys-63 polyubiquitin chains: R.M. Hofmann & C.M. Pickart; J. Biol. Chem. 276, 27936 (2001), Abstract;

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