Recombinant Human Ubiquitin Conjugating Enzyme 12, His6-Tagged

• Cat.No.: UBE2M-05H
• Product Overview: Recombinant Human Ubiquitin Conjugating Enzyme 12 produced inE.coliis a single non-glycosylated polypeptide chain containing 216 amino acids and having a molecular mass of approximately 25 kDa.

Specification

• Species: Human
• Source: E.coli
• Tag: His
• Description: UbcH12 is functional inin vitroNEDDylation reactions. It has been shown to form a thioester linkage with NEDD8 in the presence of the NEDD8 activating enzyme complex Uba3/APP-BP1. APP-BP1 binds to the amyloid precursor protein (APP) carboxy terminal domain and is important in conjunction with Uba3 and UbcH12 in driving cells through the S to M checkpoint. It was demonstrated to be the E2 responsible for the NEDDylation of the Cul-1 component of the SCF(β-TRCP) complex which is important as the E3-ligase in the ubiquitinylation of IκBα. NEDDylation of Cul-1 is essential for conjugation and processing of NF-κB p105 by SCF(β-TRCP) following phosphorylation of the complex. A dominant negative form of UbcH12, previously demonstrated to sequester NEDD8 and inhibit its conjugation, inhibits both conjugation and processing of p105, which is alleviated by wild-type UbcH12.
• Purity: >95% by SDS-PAGE and HPLC analyses.
• Formulation: Lyophilized from a 0.2μm filtered concentrated (1mg/ml) solution in 1×PBS, 1mM DTT, pH 7.5.
• Endotoxin: Less than 1EU/μg of rHuUbcH12 as determined by LAL method.
• Reconstitution: We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Reconstitute in sterile distilled water or aqueous buffer containing 0.1% BSA to a concentration of 0.1-1.0 mg/mL. Stock solutions should be apportioned into working aliquots and stored at <-20^oC. Further dilutions should be made in appropriate buffered solutions.
• Storage: This lyophilized preparation is stable at 2-8^oC, but should be kept at -20^oC for long term storage, preferably desiccated. Upon reconstitution, the preparation is stable for up to one week at 2-8^oC. For maximal stability, apportion the reconstituted preparation into working aliquots and store at -20^oC to -70^oC. Avoid repeated freeze/thaw cycles.

Gene Information

• Gene Name: UBE2M
• Synonyms: UBC12; hUbc12; UBC-RS2 ubiquitin-conjugating enzyme E2M(UBC12 homolog, yeast); ubiquitin-conjugating enzyme E2M(homologous to yeast UBC12); UBC12_HUMAN; UBE2M; NEDD8-conjugating enzyme Ubc12; EC6.3.2.-; NEDD8 protein ligase; NEDD8 carrier protein; ubiquitin carrier protein M; ubiquitin-conjugating enzyme E2M(UBC12 homolog, yeast); ubiquitin-conjugating enzyme E2M(homologous to yeast UBC12); ubiquitin-protein ligase M; yeast UBC12 homolog.
• Gene ID: 9040
• mRNA Refseq: NM_003969
• Protein Refseq: NP_003960
• MIM: 603173
• UniProt ID: P61081
• Chromosome Location: 19q13.43
• Function: ligase activity;protein binding;ribosomal S6-glutamic acid ligase activity; ubiquitin-protein ligase activity; ubiquitin-protein ligase activity.