Download 293/CRE-Luciferase Host Cell Line

293/CRE-Luciferase Host Cell Line
Technical Manual No. TM0426
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Version 10132010
Introduction…….…………………………………………………………………………….
Background………………………………………………………………………………….
Representative Data………………………………………………………………………
References ……………………………………………………………………………….
Limited Use License Agreement…………………………………………………………
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Introduction
Catalog Number: M00188
Cell Line Name: 293/CRE-Luc
Host Cell: 293
Quantity: Two vials of frozen cells (3×106 per vial)
Stability: 16 passages
Applications: Functional assays for Gs protein coupled receptors
Freeze Medium: 45% culture medium, 45% FBS, 10% DMSO
Culture Medium: DMEM, 10% FBS, 100 µg/ml Hygromycin B, 250 µg/ml G418
Mycoplasma Status: Negative
Storage: Liquid nitrogen immediately upon delivery
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Background
The G-protein-coupled receptor (GPCR) superfamily represents one of the most important classes of drug targets
in the industry. At present, about 50% of all medicines currently in the market are targeted to GPCRs. GPCRs can
be divided into several subfamilies according to the kind of Gα protein to which they couple: There are Gαs-, Gαi-,
and Gαq-coupled receptors. Gαs proteins can activate adenyl cyclase and increase the intracellular concentration
of cAMP. At GenScript, we employ both 3xCRE element and firefly luciferase as reporters to establish host cell
lines that can be used for research involving GPCRs, particularly Gs-coupled GPCRs. This cell line demonstrates
an approximately 50-fold dynamic range in the luciferase assay, providing a good screening window that is suitable
for screening GPCR agonists and antagonists.
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Representative Data
CRE Inducibility of 293/CRE/Luc Cell Line:
Unstimulated Control
Stimulated
(forskolin)
12829
699503
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54
Background subtracted luciferase
readout (RLU)
Fold induction
293/CRE-Luc
800000
54-Fold
 RLU
600000
400000
200000
0
Forskolin 10M
Cell Control
Figure: Luciferase activities were monitored with Analyst HT plate reader upon treatment with forskolin.
293/CRE-Luc cell line was treated with forskolin as indicated. Data represent means ± SEM for duplicate
samples.
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References
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Conway, S., et al., Characterisation of human melatonin MT (1) and MT (2) receptors by CRE-luciferase
reporter assay. Eur J Pharmacol, 2000. 390(1-2): p. 15-24.
2.
Klabunde, T. (2002) Drug design strategies for targeting G-protein-coupled receptors. Chembiochem. 3, 924–
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GenScript USA Inc.
860 Centennial Ave., Piscataway, NJ 08854
Tel: 732-885-9188, 732-885-9688
Fax: 732-210-0262, 732-885-5878
Email: [email protected]
Web: http://www.genscript.com
For Research Use Only.
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Limited Use License Agreement
This is a legal agreement between you (Licensee) and GenScript USA Inc. governing use of GenScript's stable cell
line products and protocols provided to licensee. By purchasing and using the stable cell line, the buyer agrees to
comply with the following terms and conditions of this label license and recognizes and agrees to such restrictions:
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The products are not transferable and will be used at the site where they were purchased. Transfer to another
site owned by buyer will be permitted only upon written request by buyer followed by subsequent written
approval by GenScript.
The purchaser cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using
this product or its components to a third party.
The products sold by GenScript are for laboratory and animal research purposes only. The products are not to
be used on humans, for consumption, or for any unlawful uses.
GenScript USA Inc. will not assert against the buyer a claim of infringement of patents owned or controlled by
GenScript USA Inc. and claiming this product based upon the manufacture, use or sale of a clinical diagnostic,
therapeutic and vaccine, or prophylactic product developed in research by the buyer in which this product or its
components has been employed, provided that neither this product nor any of its components was used in the
manufacture of such product. For information on the use of this product for other purposes, contact Marketing
Department, GenScript USA Inc., 860 Centennial Avenue, Piscataway, New Jersey 08840, U.S.A. Phone: 1-732885-9188. Fax: 1-732-210-0262. Email: [email protected].
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