Molecular Formula: N/A

CAS Number: [discontinued]

Description:
The Chemiluminescent β-Glucuronidase (GUS) Detection Kit allows for quantitative measure of GUS enzyme activity in mammalian cells, transformed plants, and purified enzyme assays through use of a chemiluminescent substrate.

Application:
The β-glucuronidase (GUS) enzyme from E. coli (EC 3.2.1.31) has been well documented to provide desirable characteristics as a marker gene in transformed plant and mammalian cells. The GUS reporter gene system has many advantages including stable expression of E. coli GUS enzyme, no interference with normal plant metabolism, and low intrinsic GUS activity in higher plants. The enzyme is also capable of tolerating amino-terminal additions, making it useful for study of plant organelle transport. Various β-glucuronic acid substrates are available for detection of GUS expression, all of which contain the sugar D-glucopyranosiduronic acid attached by glycosidic linkage to a hydroxyl group of a chromogenic, fluorogenic or chemiluminescent molecule. Upon enzymatic hydrolysis, the sugar is removed, resulting in release of the detectable molecule, and levels of gene expression can be inferred from relative color, fluorescence or light emission levels. 1,2 Dioxetane substrates emit visible light upon enzyme catalysis, providing rapid results with very low background and high intensity signal. An enhancing solution is also provided with this kit to increase light production efficiency in solution-based assays by drawing water away from the signal production site. The Chemiluminescent β-Glucuronidase Detection Kit provides all the necessary reagents, buffers, substrate, and protocols for sensitive and quantitative GUS activity assays. Please contact our technical services department for further information.

References:

• Chemiluminescent 1,2-dioxetanes. Brij P. Giri, PCT/US99/20590.
• Novel stabilized formulations for chemiluminescent assays. Brij P Giri, PCT/US01/02779.
• Single molecule detection of Alkaline Phosphatase enzyme using enhanced chemiluminescent from 1,2-dioxetanes and water-soluble, water-insoluble or partially water-soluble polymers. Brij P Giri, US Patent Application, 2002/0013250.
• Deuterium-based chemiluminescent 1,2-dioxetanes. Brij P Giri, Dinesh J Dagli, PCT application pending.
• Polymeric ammonium and or phosphonium salts having added p-electrons and higher molecular weight and enhancers for chemiluminescent systems. Brij P. Giri, US patent pending.
• Jefferson, R.A., et al., Proc. Natl. Acad. Sci. USA. 86 (1986) 8447-8451.
• Jefferson, R.A., et al., EMBO J. 6 (1987) 390-3907.
• Kosugi, S., et al., Plant Sci. 70 (1990) 130-140.
• Gallagher, S.R., et al., GUS Protocols: Using the GUS Gene as a Reporter of Gene Expression. (1992).