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Tri-ubiquitin (K63-linked)

 
BML-UW0745-0050 50 µg 149.00 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 Specification

Alternative Name:Ub3
 
MW:~25.5kDa
 
Source:Protein components produced in E. coli. Synthesised enzymatically in vitro.
 
UniProt ID:P0CG47 (UBB), P0CG48 (UBC)
 
Concentration:2.5mg/ml
 
Formulation:Liquid. In 50mM TRIS, pH 7.6, containing 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:Shipped on 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) was produced 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 (K63R-ubiquitin). The distal terminus of the resulting dimer was deblocked by treatment with yeast ubiquitin hydrolase-1 (YUH-1) followed by purification by anion exchange chromatography. The resulting Ub2 was conjugated by E1 and Ubc13-Mms2 to Asp77-ubiquitin to generate the distal-ubiquitin blocked trimer; this was deblocked with YUH-1 and purified as above to yield K63-linked tri-ubiquitin (Ub3).
 
bml-uw0745 sdspage
SDS-PAGE of tri-ubiquitin Ub3, K63-linked (Lane 2: Prod. No. BML-UW0745, 5µg), in comparison with Ub2, K63-linked (Lane 1: Prod. No. BML-UW0730, 5µg) and Ub4, K63-linked (Lane 3: Prod. No. BML-UW0715, 5µg).
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bml-uw0745 sdspage

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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For Research Use Only. Not for use in diagnostic procedures.
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