UBE2D4 (UbcH5d) [6His-tagged]


Catalogue Number
62-0015-020
Product Size
20 µg
Price £
£100
Accession Number
NP_057067
Residues Expressed
1-147
Alternate Product Size
100 µg
Certificate of Analysis Size
20 µg
Species
Human
Source
E. coli expression
Quantity
20 µg
Storage
-70°C
Concentration
1 mg/ml
Formulation
50 mM HEPES pH 7.5, 150 mM sodium chloride, 2 mM dithiothreitol, 10% glycerol
Molecular Weight
~22 kDa
Stability
12 months at -70°C; aliquot as required
Protein Sequence
Accession number: NP_057067. For full protein sequence information download the Certificate of Analysis pdf.
QA; Protein Identification
Confirmed by mass spectrometry.
QA Activity

E2-Ubiquitin Thioester Loading Assay: The activity of His-UBE2D4 was validated by loading E1 UBE1 activated ubiquitin onto the active cysteine of the His-UBE2D4 E2 enzyme via a transthiolation reaction. Incubation of the UBE1 and His-UBE2D4 enzymes in the presence of ubiquitin and ATP at 30°C was compared at two time points, T0 and T10 minutes. Sensitivity of the ubiquitin/His-UBE2D4 thioester bond to the reducing agent DTT was confirmed.


Background

The enzymes of the ubiquitylation pathway play a pivotal role in a number of cellular processes including regulated and targeted proteosomal degradation of substrate proteins. Three classes of enzymes are involved in the process of ubiquitylation; activating enzymes (E1s), conjugating enzymes (E2s) and protein ligases (E3s). UBE2D4 is a member of the E2 ubiquitin-conjugating enzyme family and the human gene was first described by Colland et al. (2004).


References

Colland F, Jacq X, Trouplin V, Mougin C, Groizeleau C, Hamburger A, Meil A, Wojcik A, Legrain P, Gauthier J (2004) Functional proteomics mapping of a human signaling pathway. Genome Res 14, 1324-32.