Ubiquitin Activating Enzyme, E1a (UBE 1a)

Ubiquitin can be covalently linked to many cellular proteins by the ubiquitination process, which targets proteins for degradation by the 26S proteasome. Three components are involved in the target protein-ubiquitin conjugation process. Ubiquitin is first activated by forming a thiolester complex with the ubiquitin-activating enzyme (UBE1 or E1). The activated ubiquitin is subsequently transferred to the ubiquitin-carrier protein E2, and then from E2 to ubiquitin ligase E3 for final delivery to the E-amino group of the target protein lysine residue (1-3). Combinatorial interactions of different E2 and E3 proteins result in substrate specificity (4). UBE1 has two isofoms: UBE1a is a nuclear protein of 117kD while UBE1b is a nuclear and cytoplasmic protein of 110kD (5). \n\nApplications:\nSuitable for use in ELISA, Western Blot, Immunoprecipitation, Immunohistochemistry, Immunocytochemistry and Immunofluorescence. Other applications not tested. \n\nRecommended Dilution: \nWestern blotting 1:1000 \nImmunoprecipitation 1:50 \nImmunohistochemistry (Paraffin) 1:175 \nIHC Protocol Citrate/TBST \nImmunofluorescence (IF-IC) 1:50 \nFlow Cytometry 1:100 \nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

U1000-09H

Size

100ul

Applications

ELISA, FC, ICC, IF, IHC, IP, WB

Hosts

Rabbit

Reactivities

Hum, Mouse, Rat

Form

Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 100ug/ml BSA and 50% glycerol.

P Type

Pab

Purity

Purified by Protein A and immunoaffinity chromatography.

References

1. Ciechanover, A. (1998) EMBO J. 17:7151-7160. 2. Hochstrasser, M. (2000) Nat. Cell Biol. 2:E153-E157. 3. Hochstrasser, M. (2000) Science 289:563-564. 4. DeSalle, L.M. and Pagano, M. (2001) FEBS Lett. 490:179-189. 5. Stephen, A.G. et al. (1996) J. Biol. Chem. 271:15608-15614.

Additional Info

Detects endogenous levels of total human UBE1a protein. Does not crossreact with UBE1b. Species Crossreactivity: Mouse and rat.