Kit for measuring cytotoxicity of a test agent

Details Kit for measuring cytotoxicity of a test agent Kit for measuring cytotoxicity of a test agent

2007-10-16

2007-10-16

Gitomer, Ralph (Department: 1657)

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving oxidoreductase

C435S975000

Reexamination Certificate

active

11087777

ABSTRACT:
Disclosed are a method and a corresponding kit for determining the cytotoxicity of a test agent. The method includes the steps of adding a pre-determined amount of the test agent to a vessel containing living cells in culture medium. The cells are then incubated for a pre-determined amount of time. To the cells and culture medium is then added a reagent mixture that is non-toxic to the living cells. In the preferred embodiment, the reagent mixture contains a solvent, a dye, an electron transfer agent, a substrate for a cytoplasmic enzyme having a half-life greater than two hours and NAD+. The contents of the vessel are then measured for production of the reduced state of the dye, the production of the reduced state of the dye being caused by a cytoplasmic enzyme specific for the substrate in the reagent mixture. In the preferred embodiment, the production of the reduced state of the dye is caused by lactate dehydrogenase released from dead and dying cells within the vessel. The kits contain components necessary for performing the method.

REFERENCES:
patent: 3107204 (1963-10-01), Brown et al.
patent: 4849347 (1989-07-01), Familletti et al.
patent: 5185450 (1993-02-01), Owen
patent: 5334508 (1994-08-01), Hoenes
patent: 5501959 (1996-03-01), Lancaster
patent: 5756527 (1998-05-01), Mjalli et al.
patent: 5766874 (1998-06-01), Miyada et al.
patent: 5858974 (1999-01-01), Little, II et al.
patent: 5912139 (1999-06-01), Iwata et al.
patent: 6057120 (2000-05-01), Heindl et al.
patent: 6350452 (2002-02-01), Riss
patent: 6811990 (2004-11-01), Corey et al.
patent: 6982152 (2006-01-01), Riss et al.
patent: 2003/0232401 (2003-12-01), Pugia et al.
patent: 1112140 (1981-11-01), None
patent: EP 0 605 370 (1994-07-01), None
patent: 0031379 (1989-02-01), None
patent: WO 01/98531 (2001-12-01), None
Amaksu et al., (1986),Tohoku J. Exp. Med., 149:343-350.
Baker et al., (1980),Microbiol., 26:248-253.
Brown et al., (1961),J. Clin. Path., 5:10-13.
Buttke et al., (1993),J. Immunol Methods, 157:233-240.
Corey et al., (1997),J. Immunol. Meth., 207:43-51.
Dunigan et al., (1995), Aqueous Soluble Tetraolium/Formazan MTS as an Indicator of NADH and NADPH Dependent Dehydrogenase Activity,BioTechniques, 19:640-649.
Evans et al., (2001),Toxicologyin Vitro, 15:579-584.
Horobin, R.W., (2001),Biotechnic&Histochemistry76(4):163-164.
Ikawa et al., (1985), Measurement of the eRatio of Primary to Total Bile Acids in Serum by Enzymatic Fluorometric Microassay and Its Clinical Significance in Patients with Liver Disease,Tohoku J. exp. Med., 145:185-195.
Kanazawa et al., (1996),J. Antibiotics, 19:229-233.
Liu, D., (1981),Bull. Environ. Contam. Toxicology, 26:145-149.
McMillan, M.K., (2002) “An Improved Resazurin Based Cytotoxicity Assay for Hepatic Cells,”Cell Biology and Toxicology, vol. 18, No. 3, pp. 157-173.
Miura et al., (1987),Analytical Biochemistry, 164:482-487.
O'Brien, J., (2000) Investigation of the Alamar Blue (Resazurin) Fluorescent Dye for the Assessment of Mammalian Cell Cytotoxicity,European J. Biochemistry, vol. 267, No. 1, pp. 5421-5426.
Rasmussen, E., (1999) “Use of Fluorescent Redox Indicators to Evaluate Cell Proliferation and Viability,” In Vitroand Molecular Toxicology, vol. 12, No. 1, pp. 47-58.
Shahangian et al., (1984),Journal of Analytical Toxicology, vol. 8, pp. 273-276.
Soyama et al., (1987),Clinica Chimica Acta, 168:259-260.
Winartasaputra et al., (1980)Clin. Chem., 26/5:613-617.

Affiliated with

Also associated with

Rating

Kit for measuring cytotoxicity of a test agent does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Kit for measuring cytotoxicity of a test agent, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Kit for measuring cytotoxicity of a test agent will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-3891550