antibody

Ubiquitin Activating Enzyme, E1a (UBE 1a)

U1000-09H

100 ul

polyclonal

domestic rabbit

nonconjugated

Ubiquitin Activating Enzyme, E1a (UBE 1a)

rabbit

Pab

Antibodies

100 ul

IgG

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 ε-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). Applications: Suitable for use in Western Blot, Immunoprecipitation, Immunohistochemistry and Immunofluorescence. Other applications not tested. Recommended Dilution: Western Blot: 1:1000 Immunoprecipitation: 1:50 Immunohistochemistry (Paraffin): 1:175, IHC Protocol Citrate/TBST Immunofluorescence (IF-IC): 1:50 Flow Cytometry: 1:100 Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 12 months at -20°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.

Hu Mo Rt

Synthetic peptide corresponding to the N-terminus of human UBE1.

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

Purified by Protein A and peptide affinity chromatography.

Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol.

Not determined

Product Type: Pab Isotype: IgG Host: rabbit Source: human Concentration: Not determined Form: Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol. Purity: Purified by Protein A and peptide affinity chromatography. Immunogen: Synthetic peptide corresponding to the N-terminus of human UBE1. Specificity: Detects endogenous levels of total human UBE1a protein. Does not crossreact with UBE1b. Species Crossreactivity: Mouse and rat. Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.

FC IF IHC IP WB

-20°C

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.

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Flow cytometric analysis of K562 cells using U1000-09H(right) compared to a nonspecific negative control antibody (left).

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Confocal immunofluorescent analysis of SK-N-MC cells using U1000-09H (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).