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InformationUbiquitin Conjuating Enzyme (E2/UBCE) ELISA Kit (Part hE2UBCE-ELISA)
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Ubiquitin Conjugating Enzyme E2/UBCE ELISA Kit
Ubiquitin Conjugating Enzyme (E2) ELISA Kit
Dynamic Assay Range: 100pg/ml - 30,000pg/ml. Sensitivity: 37 pg/ml.
Lead Time: Approximately 1.5 weeks
View Protocol
UBCE ELISA Kit Availability and Specifications
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Species |
Standard |
Assay Range |
| Human | 32ng/ml | 100pg/ml - 30ng/ml |
| Rat | 32ng/ml | 100pg/ml - 30ng/ml |
| Mouse | 32ng/ml | 100pg/ml - 30ng/ml |
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Highlights
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Intoduction
to E2/UBCE
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Ubiquitin-conjugating enzymes are known as E2 enzymes. They perform the second step of ubiquitination. The E2 enzyme receives activated ubiquitin from E1, conjugates it, then binds to E3 where it is transferred to a lysine residue on the targeted protein. The overall ubiquitination reaction causes protein degradation by proteasomes.
Ubiquitin conjugating enyme is located in the cytosol. To isolate fractions for ELISA assay, use a non-denaturing detergent lysis buffer such as Part No. ELSP-1, or similar to lyze membranes and release a soluble cytosolic fraction. The protocol associated with Part No ELSP-1 can be used to isolate cytosolic fractions for ELISA measurement.
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Methods
Overview
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The included ELISA micro-strip plate is provided pre-coated with an a monoclonal capture antibody selective to the antigen. Samples are pre-mixed with a biotin-linked detection antibody and streptavidin-HRP, which are then added to the plate. The plate is washed after a 60 min incubation period. Color reagents are added, resulting in a reaction with HRP that yields a blue colored solution. A stop solution is applied to terminate the reaction, and the solution is measured at 450nm.
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Sandwich ELISA Methods to E2/UBCE
ELISA Assay Steps
- Mix sample, biotinylated-detection antibody and streptavidin-HRP.
- Apply sample and antibody-HRP complex to ELISA plate for 60min at 37oC.Wash 5 times.
- Apply chromogens, incubate, then stop reaction.
E2/UBCE
References 1. Wiebel, Franziska F., and Wolf-H. Kunau. 1992. "The Pas2 protein essential for peroxisome biogenesis is related to ubiquitin
conjugating enzymes." Nature 359.6390, 73-76.
2. Seufert, Wolfgang, Bruce Futcher, and Stefan Jentsch.1995. "Role of a ubiquitin-conjugating enzyme in degradation of S-and M-phase cyclins." 78-81.
3. Jahngen-Hodge, Jessica, et al. 1997. "Regulation of ubiquitin-conjugating enzymes by glutathione following oxidative stress." Journal of Biological Chemistry 272.45, 28218-28226.
4. Moraes, Trevor F., et al. 2001. "Crystal structure of the human ubiquitin conjugating enzyme complex, hMms2–hUbc13." Nature Structural & Molecular Biology 8.8, 669-673.
For research only. Not for Diagnostics.
Safety: Stop solution contains acid. Avoid ingestion, skin and eye contact.
Storage: 4°C. 6 months.
Shipping. Overnight domestic delivery recommended with ice packs. Also avaialble for international delivery; please inquire for shipping cost.
 ELISA kit Fivephoton Biochemicals-80x80.jpg "UBIQUITIN LIGASE (E3) ELISA Kit (Ubiquin ligating enzyme; Part hE3UBPL-ELISA)")
