Certificate of Analysis and Data Sheet

Source:
E.Coli

Catalog No.
ENZ-274

Background:

Human UBC9 Ubiquitin-like protein SUMO-1 conjugating enzyme is also called UBE2I. rHuUBC9 Mediates the conjugation of the ubiquitin-like protein SUMO-1 to a variety of proteins including RanGAP1, IkappaBalpha, and PML without the requirement of an ubiquitin ligase (E3).

Description :

Recombinant Human ubiquitin-conjugating enzyme produced in E.Coli is a single, non-glycosylated, polypeptide chain having a molecular mass of 23 kDalton. Recombinant UBC-9 is purified by proprietary chromatographic techniques.

Physical Appearance:

Sterile Filtered solution.

Formulation:

UBC-9 ,concentration: 1 mg/ml,contains 10 mM Tris-HCl, pH 7.5, 100 mM NaCl, 100 mM imidazole, 0.5 mM PMSF, 1 mM DTT, and 10 % glycerol.

Solubility:

It is recommended to reconstitute the lyophilized UBC9 in sterile 18M?cm H2O not less than 100ug/ml, which can then be further diluted to other aqueous solutions.

Stability:

UBC9 although stable at 4^oC for 3 weeks, should be stored desiccated below -18 C. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).
Please avoid freeze-thaw cycles.

Purity:

Greater than 98.0% as determined by:
(a) Analysis by RP-HPLC.
(b) Anion-exchange FPLC.
(c) Analysis by reducing and non-reducing SDS-PAGE Silver Stained gel.

Dimers and aggregates:

Less than 1% as determined by silver-stained SDS-PAGE gel analysis.

Biological Activity:

ProSpec's IGF-I is fully biologically active when compared to standards. The ED50, calculated by the dose-dependant proliferation of murine BALB\C 3T3 cells (measured by H-thymidine uptake) is less then 1.0 ng/ml, corresponding to a specific activity of 1x10 U/mg. For most in-vitro applications, IGF-I exerts its biological activity in the concentration range of 0.2-20 ng/ml.

Endotoxin:

Less than 0.1 ng/ug (IEU/ug) of ubiquitin-conjugating enzyme

Protein content:

Protein quantitation was carried out by two independent methods:
1. UV spectroscopy at 280 nm
2. Analysis by RP-HPLC, using a standard solution of UBC-9 as a Reference Standard.

Usage:

This material is offered by ProSpec-TechnoGene for research, laboratory or further evaluation purposes.

Gene:

Name:UBE2I
Synonyms:UBC9, UBCE9

Protein synonyms/aliases:

Ubiquitin-conjugating enzyme E2 I (EC 6.3.2.19)
    (Ubiquitin-proteinligase I)
    (Ubiquitin carrier protein I)
    (SUMO-1-protein ligase)
    (SUMO-1-conjugating enzyme)
    (Ubiquitin carrier protein 9)
    (p18).

Protein Family:

Belongs to the ubiquitin-conjugating enzyme family.

Protein Domains:

Domains:
•IPR000608   Ubiquitin-conjugating enzyme, E2

Protein links:

Precursor- Protein structure and amino acid sequence:

Latest Publications:

1. Cloning, characterization and subcellular localization of a gene encoding a human Ubiquitin-conjugating enzyme (E2) homologous to the Arabidopsis thaliana UBC-16 gene product.
Front Biosci 2006 May 1;11:1500-7
2. Mechanistic insight into the allosteric activation of a ubiquitin-conjugating enzyme by RING-type ubiquitin ligases.
Proc Natl Acad Sci U S A 2005 Dec 27;102(52):18890-5
3. The ubiquitin-protein ligases Nedd4 and Nedd4-2 show similar ubiquitin-conjugating enzyme specificities.
Int J Biochem Cell Biol 2006 Mar;38(3):472-9
4. Purification and activity assays for Ubc9, the ubiquitin-conjugating enzyme for the small ubiquitin-like modifier SUMO.
Methods Enzymol 2005;398:74-87
5. Expression, purification, and structural analysis of (HIS)UBE2G2 (human ubiquitin-conjugating enzyme).
Protein Expr Purif 2006 Feb;45(2):324-8
6. NMR assignment of human ubiquitin conjugating enzyme Ubc7.
J Biomol NMR 2005 Aug;32(4):340

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中華民國95年06月06日更新