UBE2G1 (UBC7), His-Tag Recombinant

Catalog #

79372

$325 *

Size: 20 µg

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Description

Recombinant full length human UBE2G1 (ubiquitin-conjugation enzyme E2 G1). This construct contains an N-terminal His-tag. The recombinant protein was affinity purified and is active.

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Synonyms

UBC7, E217K, UBE2G

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Product Data Gallery

4-20% SDS-PAGE Coomassie Staining

Specific Activity

Product Info

Storage and Usage

Citations

Species

Human

Construct

UBE2G1 (His-Full length)

Host Species/Expression System

E. coli

Purity

≥90%

Format

Aqueous buffer solution

Formulation

50 mM sodium phosphate, pH 7.0, 300 mM NaCl, 150 mM imidazole, 0.1 mM PMSF, 0.25 mM DTT, 25% glycerol.

21 kDa

Specific Activity

11 pmol/min/µg

Genbank #

NM_003342

UniProt #

P62253

Tag(s)

N-terminal His-tag

Background

UBE2G1 (UBC7) or ubiquitin-conjugating enzyme E2G 1 is a member of the E2 ubiquitin-conjugating enzyme family and catalyzes the covalent attachment of ubiquitin to other proteins which are involved in degradation of muscle-specific proteins. UBC7 is highly expressed in skeletal muscle (1). UBC7 is involved in the selective ubiquitylation of different substrates by the same E3 complex (2).

References

1. Watanabe, T.K., et.al., Cytogenet. Cell Genet. 1996; 74: 146-148.

2. Shibata, E., et al., Mol Cvell Biol. 2011 Aug.

Storage/Stability

At least 6 months at –80°C.

Assay Conditions

UBE2G1 activity was measured by using the wild type ubiquitin protein diluted in Ubiquitination Buffer [(40 mM Tris (pH 7.5), 20 mM MgCl₂, 0.1 mg/ml BSA and 0.5 mM DTT] to a working concentration of 170 ng/µl (20 µM), in AMP-Glo™ based assay (Promega #V5011). Reaction was initiated by mixing 5 µl of a solution of 2x the final concentrations of UBE2G1 with 5 µl of a 2x Reaction Cocktail (1 µl of stock solution of ubiquitin, 15 ng/ µl UBA1 + 40 ng/µl BIRC3 + 50 µM ATP).

The reaction was initiated by mixing 5 µl of 2x UBE2G1 protein dilutions with 5 µl of 2x Reaction Cocktail for 60 minutes at 37°C. After the plate was equilibrated to Room Temperature (RT), 10 µl of AMP-Glo™ Reagent I was added, mixed for 1-2 minutes and incubated at RT for 60 minutes. AMP-Glo™ Reagent II was diluted 100-fold with Kinase-Glo™ One Solution, and 20 µl were added to each well, mixed for 1-2 minutes and incubated at RT for 30 minutes. The plate was read in a GloMax plate reader (Promega # E7031). The Enzyme Specific Activity was calculated as follows: [ (AMP concentration in µM) * (Reaction Volume in µl)] / Reaction time in min) *(Enzyme amount in mg)] x 10^-3. A blank can be determined from a “no substrate” sample by replacing the enzyme solution with an equal volume of Ubiquitination Buffer.

Instructions for Use

Thaw on ice and gently mix prior to use. DO NOT VORTEX. Perform a quick spin before opening. Aliquot into small volumes and flash freeze for long term storage. Avoid multiple freeze/thaw cycles.

Applications

Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.

Shipping Temperature

-80°C

Warnings

Avoid freeze/thaw cycles

Scientific Category

Ubiquitination

Regulatory Status

Research Use Only

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Datasheets/Protocols

79273 Lot 231025

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UBE2G1 (UBC7), His-Tag Recombinant

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