| Product Includes | Product # | Quantity | Color |
|---|---|---|---|
| FastScan™ ELISA Microwell Strip Plate, 96 Well | 53257 | 96 tests | |
| Cyclin E1 Mouse Capture mAb | 62903 | 1 ea | Green (Lyophilized) |
| Cyclin E1 Rabbit HRP-linked mAb | 90179 | 1 ea | Red (Lyophilized) |
| FastScan™ ELISA Capture Antibody Diluent | 16076 | 3 ml | Green |
| FastScan™ ELISA HRP Antibody Diluent | 28120 | 3 ml | |
| TMB Substrate | 7004 | 11 ml | |
| STOP Solution | 7002 | 11 ml | |
| Sealing Tape | 54503 | 1 ea | |
| ELISA Wash Buffer (20X) | 9801 | 25 ml | |
| FastScan™ ELISA Cell Extraction Buffer (5X) | 69905 | 10 ml | |
| FastScan™ ELISA Cell Extraction Enhancer Solution (50X) | 25243 | 1 ml | |
| FastScan™ ELISA Kit #45829 Positive Control Type 2 | 24826 | 1 ea |
*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.
Description
The FastScan™ Total Cyclin E1 ELISA Kit is a sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of cyclin E1. To perform the assay, sample is incubated with a capture antibody conjugated with a proprietary tag and a second detection antibody linked to HRP, forming a sandwich with cyclin E1 in solution. This entire complex is immobilized to the plate via an anti-tag antibody. The wells are then washed to remove unbound material. TMB is then added. The magnitude of observed signal is proportional to the quantity of cyclin E1.
*Antibodies in kit are custom formulations specific to kit.
IMPORTANT: This FastScan™ ELISA Kit requires 4 washes at Step 6 of the protocol.
Specificity/Sensitivity
Background
Cyclin E1 and cyclin E2 can associate with and activate CDK2 (1). Upon DNA damage, upregulation/activation of the CDK inhibitors p21 Waf1/Cip1 and p27 Kip1 prevent cyclin E/CDK2 activation, resulting in G1/S arrest. When conditions are favorable for cell cycle progression, cyclin D/CDK4/6 phosphorylates Rb and is thought to reduce the activity of p21 Waf1/Cip1 and p27 Kip1, allowing subsequent activation of cyclin E/CDK2 (1,2). Cyclin E/CDK2 further phosphorylates Rb to allow progression into S-phase, where cyclin E/CDK2 is thought to phosphorylate and activate multiple proteins involved in DNA synthesis (2,3). Turnover of cyclin E is largely controlled by phosphorylation that results in SCFFbw7-mediated ubiquitination and proteasome-dependent degradation (4,5). Cyclin E1 is phosphorylated at multiple sites in vivo including Thr62, Ser88, Ser72, Thr380, and Ser384, and is controlled by at least two kinases, CDK2 and GSK-3 (6,7).
- Lauper, N. et al. (1998) Oncogene 17, 2637-43.
- Lundberg, A.S. and Weinberg, R.A. (1998) Mol Cell Biol 18, 753-61.
- Ewen, M.E. (2000) Genes Dev 14, 2265-70.
- Won, K.A. and Reed, S.I. (1996) EMBO J 15, 4182-93.
- Koepp, D.M. et al. (2001) Science 294, 173-7.
- Welcker, M. et al. (2003) Mol Cell 12, 381-92.
- Ye, X. et al. (2004) J Biol Chem 279, 50110-9.
Background References
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
Trademarks and Patents
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