Life & Death Assay Kit

BV-K501-100

50 determinations

Distinguishing between live and dead cells is very important for investigation of growth control and cell death. The Live-Dead Cell Staining Kit provides ready-to-use staining reagents to conveniently discriminate between live and dead cells. The kit utilizes Live-Dye^TM, a cell-permeable green fluorescent dye (Ex/Em = 488/518 nm), to stain live cells. Dead cells can be easily stained by propidium iodide (PI), a cell non-permeable red fluorescent dye (Ex/Em = 488/615). Stained live and dead cells can be visualized by fluorescence microscopy using a band-pass filter (detects FITC and rhodamine). Ready-to-use.

Cell Counting Kit-8 [CCKi-8]

ALX-850-039

Quantity: 500 tests / 5ml; 2500 tests / 5 x 5ml.

Kit/Set Contains: 5ml vial (5mM WST-8, 0.2mM 1-methoxy-PMS, 150mM NaCl).

Formulation: 5mM WST-8, 0.2mM 1-methoxy-PMS, 150mM NaCl.

Product Description:

Colorimetric kit for simple and accurate cell proliferation and cytotoxicity assays.

Advantages:

One-step, ready-to-use solution, no radioisotopes, no solubilization step, high sensitivity, correlates with the [3H]-Thymidine Incorporation Assay (see Figure).

Principle: Employs the tetrazolium salt WST-8 (2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium . monosodium salt), that produces a highly water soluble formazan dye upon biochemical reduction in the presence of an electron carrier, 1-methoxy-PMS. The amount of the yellow colored formazan dye generated by dehydrogenases in cells is directly proportional to the number of viable cells in a culture medium.

CCKi-8 is supplied as a ready-to-use solution. Furthermore, no radioisotopes or organic solvents are required. It can be added directly to the cell media for fast, high-throughput screening without a solubilization process. The user obtains highly reproducible and accurate results.

Product Specific Literature References:

M. Ishiyama, et al.; Biol. Pharm. Bull. 19, 1518 (1996)

J.S. Elder; Pediatr. Clin. North Am. 44, 1299 (1997)

M. Ishiyama, et al.; Talanta 44, 1299 (1997) / I. Isobe, et al.; Neurosci. Lett. 266, 129 (1999)

H. Tominaga, et al.; Anal. Commun. 36, 47 (1999) / M. Matsuoka, et al.; Biochem. Pharmacol. 59, 1573 (2000)

Cell Counting Kit-F [CCKi-F] [500 tests]

ALX-850-245-KI01

Fluorometric kit for simple and accurate cell proliferation and cytotoxicity assays.

Advantages: No radioisotopes, more sensitive than the colorimetric assay, short staining reaction time, no solubilization step. Principle: The amount of a fluorescent dye, calcein, produced from calcein-AM (3',6'-Di(O-acetyl)-2',7'-bis[N,N-bis-(carboxymethyl)aminomethyl]fluorescein . tetraacetoxymethyl ester) by esterases in cells is directly proportional to the number of viable cells in a culture medium. Since calcein-AM is highly lipophilic due to the acetoxymethyl groups, it can rapidly permeate into the cytoplasma through the cell membrane. The CCKi-F assay does not require any radioisotopes or a solubilization procedure. Therefore it allows the user to obtain highly reproducible and accurate results. The detection range of the number of viable cells by CCKi-F is from <50 to at least 25,000 cells. CCKi-F correlates with the [³H]-thymidine incorporation assay and can therefore substitute for it.

ApoSENSOR ATP Depletion Assay Kit

ALX-850-247

Kit/Set Contains: Nucleotide Releasing Buffer, ATP Monitoring Enzyme, Enzyme Reconstitution Buffer, ATP.

Product Description:

The assay utilizes bioluminescent detection of the ATP level for a rapid screening of apoptosis and cell proliferation simultaneously in mammalian cells. The assay utilizes the enzyme luciferase to catalyze the formation of light from ATP and luciferin. The light can be measured using a luminometer or Beta Counter. The assay can be fully automatic for high throughput (10 seconds/sample) and is highly sensitive (detects 10-100 mammalian cells/well).

Cell Proliferation Assay Fluorescent DNA Stains ABSOLUTE-S Kit

ALX-850-043-KI01

Description of Kit

The kit utilizes the SBIP (strand break induced photolysis) methodology in wich cells are irradiated with UV light to induce strand breaks. That does not require DNA denaturation and, therefore, is applicable in studies where preservation of antigens of other features of the cells is desired. The kit provides the most powerful technique for studies of both cell proliferation and cell death.

The Phoenix Flow Systems, Inc. ABSOLUTE-STM Kit is a two color staining method for measuring cell proliferation by multiparameter analysis of DNA replication and cellular DNA content/cell cycle position by flow or image cytometry (6). The kit contains instructions and all reagents which include BrdUrd PhotolyteTM and Photolyte EnhancerTM solutions; washing, reaction and rinsing buffers for processing individual steps in the assay; terminal deoxynucleotidyl transferase enzyme (TdT), bromodeoxyuridine triphosphate (Br-dUTP), fluorescein labeled antiBrdUrd antibody (F~PRB-1) for labeling DNA breaks and propidium iodide/RNase A solution for counter staining the total DNA. In addition, two types of control cells which address the two variables of this cell proliferation assay are included: "Reaction" and "Photolysis". The "Reaction" control cells are fixed cultured cells which have BrdU incorporated into them and have been exposed to UV light to induce the breaks in the DNA. These cells should be stained without any other treatment. The "Photolysis" control cells are fixed cultured cells which have been incubated with BrdU but have not been exposed to UV light. These cells should be exposed to UV light using the same light source and exposure time as the experimental cells.

Quantity: 50 assays. Contains all the reagents necessary, including positive and negative control cells.

Product Specific Literature References:

Detection of 5-bromo-2-deoxyuridine incorporated into DNA by labeling strand breaks induced by photolysis (SBIP): X. Li, et al.; Int. J. Oncol. 4, 1157 (1994)

Detection of apoptosis and DNA replication by differential labeling of DNA strand breaks with fluorochromes of different color: X. Li, et al.; Exp. Cell. Res. 222, 28 (1996)

Cell Proliferation Photolysis Control Cells

PFS-ASNC12

Cell Proliferation Reaction Control Cells

PFS-ASPC11

Lactate Dehydrogenase (LDH) Cytotoxicity Detection Kit (TaKaRa)

TAK-MK401

For the measurement (2000 tests) of lactate dehydrogenase (LDH) (a stable cytoplasmic enzyme present in most cells), which is released into cell culture supernatant upon damage of the cytoplasmic membrane.

Description
Cell death is assayed by the quantification of plasma membrane damage. Several standards methods for quantification of cellular viability were developed from the need for the sensitive, reliable and automated methods for precise determination of cell death.
Widely used standard methods are based on the uptake or exclusion of dyes such as trypan blue or eosin Y. These methods have the following disadvantages; 1) can not process large numbers of sample, 2) can not quantitate dead cells which may have damaged.
The second standard methods are based on the measurement of the amount of released radioactive isotopes or fluorescence dyes from the target cells which are prelabeled with those substances. The disadvantages of these methods are 1) need to prelabel target cells before assay, 2) prelabeled target cells release most labels spontaneously.
The third standard methods are based on the measurement of cytoplasmic enzyme activity released from damaged cells. The amount of enzyme activity correlates to the proportion of the damaged cells. Alkaline and acid phosphatase, or glutamic-oxalocetic transaminase (GOT), or glutamicpyruvic transaminase (GPT) have been conventionally used for the methods. However, this method is not widely used because the low amount of these enzymes are present in many cells and because the kinetic assays to quantitate these enzymes are elaborate.
Lactate dehydrogenase (LDH) is a stable cytoplasmic enzyme which is present in most cells. It is released into the cell culture supernatant upon damage of the cytoplasmic membrane. Takara's LDH Cytotoxicity Detection Kit allows simple measurement of LDH activity released from the damaged cells into the supernatant.

Principle

The culture supernatant is incubated with the reaction mixture supplied in the kit. Then, the LDH activity is measured in an enzymatic test. (See Figure)

1st step) NAD⁺ is reduced to NADH/H⁺ by the LDH-catalyzed conversion of lactate to pyruvate.

2nd step) The catalyst (diaphorase) transfers H/H⁺ of NADH/H⁺ to the tetrazolium salt INT which is reduced to formazan. This leads to color change from pale yellow to red.

Figure Principle of measurement

Features
Kit components Bottle 1 (blue cap) x 5:: Catalyst (diaphorase/ NAD⁺ )
Bottle 2 (red cap) x 5:: Dye solution (45 ml)
(containing iodotetrazolium chloride (INT) and sodium lactate)

Storage
20C

1) Cook, J. A. & Mitchell, J. B. (1989) Anal. Biochem. 179, 1-7.
2) Yuhas, J. M., Toya, R. E. & Pazmino, N. H. (1974) J. Nal. Cancer Inst. 53,465-468.
3) Parks, D. R. et al. (1979) Proc. Nail. Acad. Sci USA 76, 1979.
4) Jones, K. H. & Senll, J. A. (1985) J. Hislochem. Cytochem. 33, 77-79.
5) Oldham, R. K. et al. (1977) J. Nal. Cancer Inst. 58,1061-1067.
6) Leibold, W. & Bridge, S. (1979) Z. Immunilalsforschung(Immunobiology) 155, 287-311.
7) Kolber, M. A. et al. (1988) J. Immunol. Meth. 108, 255-264.
8) Danks, A. M. et al. (1992) Molecular Brain Research 16, 168-172.
9) Szekeres, J., Pacsa, A. S. & Pejlsik, B. (1981) J. Immun. Meth. 40, 151-154.
10) Masanel, J., Gomez-Lechon, M. J. & Castell, J. V. (1988) Toxic, in Vitro 2, 275-282.
11) Martin, A. & Clynes, M. (1991) In Vitro Cell Dev. Biol. 27A, 183-184.
12) Decker, T. & Lohmann-Mallhes, M. L. (1988) J. Immun. Meth. 15, 61-69.
13) Korzeniewski, C. & Callewaert, D. M. (1983) J. Immun. Meth. 64, 313-320.
14) Dubar, V. et al. (1993) Exp. Lung Res. 19, 345-359.
15) Kondo, T. et al. (1993) Toxic, in Vitro 7, 61-67.
16) Murphy, E. J., Roberts, E. & Horrocks, L. A. (1993) Neuroscience 55, 597-605.
17) Courjaull, F. et al. (1993) Arch. Toxicol. 67, 338-346.
18) Shrivaslava, R. et al. (1992) Cell Biology and Toxicology 8, 157-170.
19) Gelderblom, W. C. A. et al. (1993) Fd. Chem. Toxic. 31, 407-414.
20) Thomas, J. P., Geiger, P. G. & Girolli, A. W. (1993) Journal ol Lipid Research 34, 479-490.
21) Sasaki T. et al. (1992) Toxic, in Vitro 6, 451-457.
22) Goergen, J. L., Marc, A. & Engasser, J. M. (1993) Cytotechnology 11, 189-195.
23) Legrand, C. et al. (1992) J. Biotechnol. 25, 231-243.
24) Racher, A. J., Looby, D. & Grilfiths, J. B. (1990) Cytotechnology 3, 301-307.

Premix WST-1 Cell Proliferation Assay Kit

TAK-MK400-KI01

Description
In the life science it is very important to measure the cell proliferation or the cell viability.
Some standrard methods were developed for satisfying the need of sensitive, accurate, speedy, and simple method. In these methods DNA synthesis is detected by the measurement of RI-labeled nucleoside uptaken into DNA , as DNA synthesis is accompanied with the cell proliferation or the cell viability.
Proliferation assays have become available for analyzing the number of viable cells are measured by the detection of cleaved tetrazolium salts that added into a medium. Wash and collection of cells are not required, and all procedure, from the culture in small scale to the data analysis with ELISA reader, can be carried out in the same microtiter plate.
The PreMix WST-1 enables to measure the cell proliferation and cell viability with colorimetric assay, and bases on the cleavage of tetrazolium salts by mitochondrial dehydrogenase in viable cells. This product takes the place of RI-labeled nucleoside, and provides non-RI method for the analysis of cell proliferation or cell viability.

Principle
Tetrazolium salt (WST-1) is cleaved to soluble formazan dye by the succinate-tetrazolium reductase (EC 1.3.99.1), which exists in mitochondrial respiratory chain and is active only in viable cells. Total activity of this mitochondrial dehydrogenase in a sample rises with the increase of viable cells. As the increase of enzyme activity leads to an increase of the production of formazan dye, the quantity of formazan dye is related directly with the number of metabolically active cells in the medium. The formazan dye formed by metabolically active cell can be quantitated by measuring its absorbance by ELISA reader. The absorbance of formazan dye solution is in direct proportion to the number of viable cells. (Figure 1) This product is also designed for non-radioactive and spectrophotometric quantification of cell growth and viability in proliferation and chemosensitivity assay.

Intended Usage

The measurement of cell proliferation which is sensitive to growth factors, cytokines, mitogens, and nutrients
The analysis of cytotoxic and cytostatic compounds such as anticancer drugs, etc.
The evaluation of physiological mediator and antibodies which inhibit cell growth.

(EC = electron coupling reagent, RS = mitochondrial succinate-tetrazolium-reductase system)

Figure 1 Cleavage of the tetrazolium salt (WST-1) to formazan

Features
Kit components
25 ml x 1 bottle of PreMix WST-1: The PreMix WST-1 is a ready-to-use solution containing WST-1 and an electron coupling reagent, diluted in phosphate buffered saline, sterile.
Storage

If the precipitate is observed when dissolving this reagent, redissolve any precipitate by warming at 37C for several minutes with shaking gently. Once thawed, it can be stored at 4C, protected from light, for several weeks. For longer storage, it is recommended to store in aliquots at 20C. Do not repeat freeze-thaw cycle.

References:

1) Cook, J. A. & Mitchell, J. B. (1989) Anal. Biochem 179, 1-7.
2) Mosmann, T. (1983) J. Immunol. Mthods 65, 55-63.
3) Carmichael, J. et al. (1987) Cancer Res. 47, 936-942.
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5) Scudiero, D. A. et al. (1988) Cancer Res. 48, 4827-4833.
6) Weislow, O. S. et al. (1989) J. Natl. Cancer. Inst. 81, 577-586.
7) Roehm, N. W. et al. (1991) J. Immunol. Methods 142, 257-265.
8) Cory, A. H. et al. (1991) Cancer Commun. 3, 207-212.
9) Slater, T. F., Sawyer, B. & Strault, U. (1963) Biochim. Biophys. Acta 77, 383-393.

Quick Cell Proliferation Assay Kit

BV-K301-500 500 assays

BV-K301-2500 2500 assays

The Quick Cell Proliferation Assay Kit provides all reagents and detailed instructions for a fast and sensitive quantification of cell proliferation and viability. The assay is based on the cleavage of the tetrazolium salt WST-1 to formazan by cellular mitochondrial dehydrogenases. Expansion in the number of viable cells resulted in an increase in the activity of the mitochondrial dehydrogenases, which leads to the increase in the amount of formazan dye formed. The formazan dye produced by viable cells can be quantified by multi-well spectrophotometer (microtiter plate reader) by measuring the absorbance of the dye solution at 440 nm. The assay can be used for the measurement of cell proliferation in response to growth factors, cytokines, mitogens,

and nutrients, etc. It can also be used for the analysis of cytotoxic compounds like anticancer drugs and many other toxic agents and pharmaceutical compounds. The new method is so simple, requiring no washing, no harvesting, and no solubilization steps, and is faster and more sensitive than MTT, XTT, or MTS-based assays. The entire assay can be performed in a microtiter plate.

Kit Contents:

WST-1 Reagent (lyophilized)

Electro Coupling Solution (ECS)

Bioluminescence Cytotoxicity Assay Kit

BV-K312-500

500 assays

Cell death or cytotoxicity is classically evaluated by the quantification of plasma membrane damage. Adenylate kinase (AK) is a ubiquitous protein present in all eukaryotic and prokaryotic cells and rapidly release into culture medium upon damage of the plasma membrane. The Bioluminescence Cytotoxicity Assay Kit is based on the measurement of AK in a simple one-step procedure involving two chemical reactions. The first reaction is the conversion of ADP to ATP by adenylate kinase that was released from the damaged cells. The second reaction

utilizes luciferase to catalyze the formation of light from ATP and luciferin, and the light can be measured using a luminometer or beta counter. The assay is highly sensitive and can be fully automatic for high throughput.

Kit Contents:

AK Detection Reagent (Lyophilized) 5 vials

AK Assay Buffer 50 ml

Senescence Detection Kit

BV-K320-250

250 Stainings.

Designed to histochemically detect specific senescence marker in distinct pH in cultured cells and tissue sections. The specific marker is only present in senescent cells (not found in presenescent/quiescent/immortal cells.

Senescence is thought to be a tumor suppressive mechanism and an underlying cause of aging. Senescence represents an arrested state in which the cells remain viable, but not stimulated to divide by serum or passage in culture. Senescent cells display increase of cell size, senescence-associated expression of b-galactosidase (SA-b-Gal) activity, and altered patterns of gene expression. The Senescence detection kit is designed to histochemically detect SA-b-Gal activity in cultured cells and tissue sections, a known characteristic of senescent cells. The SA-b-Gal is present only in senescent cells and is not found in presenescent, quiescent or immortal cells.

Apoptotic DNA Ladder Detection

DNA Laddering Kit [Content: 24 reactions]

CAY-660990

Quick Apoptotic DNA Ladder Detection Kit (Kit content: 50 tests) (BioVision)

ALX-850-242