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Ubiquitin (human), (fluorescein labeled)

BML-UW1240-0100 100 µg 363.00 USD
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Product Details

Source:Produced in E. coli.
UniProt ID:P0CG47 (UBB), P0CG48 (UBC), P62979 (RPS27A), P62987 (UBA52)
Purity:≥95% (SDS-PAGE)
Purity Detail:Purified by multi-step chromatography.
Application Notes:For use in ubiquitinylation studies.
Reconstitution:Reconstitute in minimum volume of DMSO (5-10µL) followed by dilution to required concentration with aqueous buffer of choice (e.g. 1mg/mL in PBS).
Shipping:Blue Ice
Short Term Storage:-20°C
Long Term Storage:-20°C
Use/Stability:Stable for at least 1 year after receipt when stored at -20°C.  Reagent is light sensitive; store in the dark or in amber vials. Stable to freeze/thaw but this should be minimized wherever possible.
Handling:Protect from light.
Scientific Background:The covalent attachment of ubiquitin to proteins (ubiquitinylation) and their subsequent proteasomal degradation plays a fundamental role in the regulation of cellular function through biological events involving cell cycle, differentiation, immune responses, DNA repair, chromatin structure, and apoptosis.
Technical Info/Product Notes:Fluorescein labelled ubiquitin is a useful reagent for studying ubiquitinylation in vitro, where it represents a readily detectable alternative to radio-iodinated ubiquitin. Fluorescein labelled ubiquitin is activated by ubiquitin E1, is a competent substrate for known ubiquitin E2/E3 enzymes (thioester bond formation) and can be conjugated to ubiquitin target proteins (isopeptide bond formation).
This product contains a number of multiply fluorescein-labelled ubiquitin species, with [fluorescein]2-4ubiquitin being the predominant species.

Typical thioester assay set-up: 2.5µM Fluorescein-Ub (BML-UW1240), 100nM Ub-E1 (BML-UW9410), 2.5µM E2 (e.g. His6-UbcH5a, BML-UW9050), 1U inorganic pyrophosphatase, 5mM Mg-ATP in 20mM TRIS-HCl, pH 7.5, 1mM DTT.  Incubate at 37ºC for 30-60 mins. Analyse by SDS-PAGE followed by fluorescence detection.
Regulatory Status:RUO - Research Use Only

Product Literature References

Mechanisms underlying ubiquitination: C.M. Pickart; Annu. Rev. Biochem. 70, 503 (2001), Abstract;
The ubiquitin-proteasome system and endocytosis: G.J. Strous & R. Govers; J. Cell Sci. 112 (Pt 10), 1417 (1999), Abstract;
The ubiquitin system: A. Hershko et al.; Annu. Rev. Biochem. 67, 425 (1998), Abstract;
Pathways of ubiquitin conjugation: A.L. Haas & T.J. Siepmann; FASEB J. 11, 1257 (1997), Abstract;

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