Revision 7

#45829

Store at +4C

FastScan Total Cyclin E1 ELISA Kit

1 Kit

(96 assays)

Species Cross Reactivity:
H

UniProt ID:
#P24864

Entrez-Gene Id:
#898

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Orders:

877-616-CELL (2355)

orders@cellsignal.com

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Product Includes Product # Quantity Color
FastScan™ ELISA Microwell Strip Plate, 96 Well5325796 tests
Cyclin E1 Mouse Capture mAb629031 eaGreen (Lyophilized)
Cyclin E1 Rabbit HRP-linked mAb901791 eaRed (Lyophilized)
FastScan™ ELISA Capture Antibody Diluent160763 mlGreen
FastScan™ ELISA HRP Antibody Diluent281203 ml
TMB Substrate700411 ml
STOP Solution700211 ml
Sealing Tape545031 ea
ELISA Wash Buffer (20X)980125 ml
FastScan™ ELISA Cell Extraction Buffer (5X)6990510 ml
FastScan™ ELISA Cell Extraction Enhancer Solution (50X)252431 ml
FastScan™ ELISA Kit #45829 Positive Control Type 2248261 ea

Kit contents scale proportionally with size, except sealing tape.
Example: The V1 kit contains 5X the listed quantities above, but will exclude the sealing tape.

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

The FastScan™ Total Cyclin E1 ELISA Kit detects endogenous levels of cyclin E1, as shown in Figure 1. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

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).

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

FastScan™ ELISA is a registered trademark of Cell Signaling Technology, Inc.

U.S. Patents 9,086,407, 9,261,500, and 9,476,874, foreign equivalents, and child patents deriving therefrom.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) orders@cellsignal.com Support: 877-678-TECH (8324) info@cellsignal.com Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 7

Cell Signaling Technology Logo
Figure 1. Cyclin E1 protein is highly expressed in U-2 OS cells as compared to Hep G2 cells. The relationship between lysate protein concentration from U-2 OS and Hep G2 cells and the absorbance at 450 nm using the FastScan™ Total Cyclin E1 ELISA Kit #45829 is shown in the upper figure. The corresponding western blot using cyclin E1 antibody is shown in the lower figure. U-2 OS and Hep G2 cells were grown to confluence at 37°C and then lysed.
ELISA Image 1: FastScan<sup>™</sup> Total Cyclin E1 ELISA Kit
Orders: 877-616-CELL (2355) orders@cellsignal.com Support: 877-678-TECH (8324) info@cellsignal.com Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.