Home Shop Ubiquitin (human), (fluorescein labeled)

Ubiquitin (human), (fluorescein labeled)

BML-UW1240

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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.

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Product Details

Application Notes	For use in ubiquitinylation studies.
Formulation	Lyophilized.
MW	~9.5kDa
Purity	≥95% (SDS-PAGE)
Purity Detail	Purified by multi-step chromatography.
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).
Source	Produced in E. coli.
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.
UniProt ID	P0CG47 (UBB), P0CG48 (UBC), P62979 (RPS27A), P62987 (UBA52)

Handling & Storage

Use/Stability	As indicated on product label or CoA when stored as recommended. 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.
Short Term Storage	-20°C
Long Term Storage	-20°C
Shipping	Blue Ice

Regulatory Status	RUO – Research Use Only

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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Ubiquitin (human), (fluorescein labeled)

BML-UW1240

Product Details

Handling & Storage

Datasheet, Manuals, SDS & CofA

Manuals And Inserts

Certificate of Analysis

SDS

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