There is great interest in improved methods to quantitate cell proliferation and survival,
which are important for applications in cell biology, toxicology, drug screening, and many
other biosciences. All current approaches on the market in use for several decades have a variety
of disadvantages, including non-linearity, high background, and cumbersome, costly, and time-consuming protocols.
CellCountEZ® is a tissue culture media-based assay that can measure metabolically active live
cells and quantify cell death caused by radiation, chemotherapeutics, or toxins. The compound in
CellCountEZ® is superior to existing compounds used for quantifying cell
proliferation and survival because it is readily soluble, membrane permeable, and converted by live
cells intracellularly before transport into the extracellular culture media. Cell media is used in
the assay, avoiding the need to lyse cells and thereby saving time and cost while preserving the ability
to perform other cellular tests in the same culture system. Thus, CellCountEZ® offers
many advantages that make it superior to common existing methods for quantifying cell growth and survival.
CellCountEZ® Kit Reagents
Reagent 1 - Store at 2 - 25° C
Reagent 2 - Store at 2 - 8° C
Reagent 3 - Store at 2 - 8° C
Approximate uses: 1000 assays using a 96-well plate. Custom designed kits are also available for 6 and 24-well plates. Please contact Rockland at email@example.com for more information.
Preparation of Reagent 4: Transfer 1.2 ml of Reagent 2 into Reagent 1 in the amber microfuge tube and vortex for 15 to 30 seconds. Bring Reagent 4 to room temperature before preparing Reagent 5.
Preparation of Reagent 5: Five minutes before the assay, add 100 µl of room temperature Reagent 4 to 900 µl (1:9) of your growth medium with no more than 15% fetal bovine serum in a new microfuge tube and mix it gently with a micropipette. Avoid air bubbles.
Eye, skin and respiratory irritants are contained in this kit. Do not ingest or inhale. Utilize standard laboratory safety procedures when handling these reagents. Please refer to the MSDS online for more information. FOR LIFE SCIENCE RESEARCH USE ONLY.
CellCountEZ® TPS Assay
Add 10 µl of Reagent 5 to each well of a 96-well plate. Incubate for 2 hours at 37° C in a humidified CO2 incubator.
Add 30 µl of Reagent 3 to each well of the above plate. Incubate for 1-2 minutes at room temperature. Read at 412 nm with a reference wavelength of 650 nm in a microtiter plate reader.
Transfer 80 µl medium from Step 1 into another 96-well plate. Add 30 µl of Reagent 3 to each well of that plate. Incubate for 1-2 minutes at room temperature. Read at 412 nm with a reference wavelength of 650 nm in a microtiter plate reader.
This assay gives a linear response for cells (0; 1,000; 5,000; 10,000; 20,000; 40,000) plated in 100 µl growth medium with up to 15% fetal bovine serum in a 96-well plate and measured 20 hours after plating.
Drug toxicity could be measured for up to 4 days after treatment with drugs using this assay for 5,000 cells plated in 100 µl growth medium with up to 15% fetal bovine serum in a 96-well plate.
- Li J, Zhang D, Ward KM, Prendergast GC and Ayene IS. Hydroxyethyl disulfide as an efficient metabolic assay for cell viability in vitro. Toxicol. In Vitro, 26(4):603-12.
- Ayene IS, Biaglow JE, Kachur AV, Stamato TD and Koch CJ (2008). Mutation in G6PD gene leads to loss of cellular control of protein glutathionylation: Mechanism and Implication. J. Cell Biochem. 103:123-135.
- Ayene IS, Stamato TD, Mauldin SK, Biaglow JE, Tuttle SW, Jenkins SF and Koch CJ (2002). Mutation in the glucose-6-phosphate dehydrogenase gene leads to inactivation of Ku DNA end binding during oxidative stress. J. Biol. Chem. 277:9929-9935.
To Place an Order
By Phone: 484-476-3474
By Fax: 484-476-8533
By Email: firstname.lastname@example.org
LIMR Development, Inc.
Attn: J. Wynne
100 East Lancaster Avenue
Wynnewood, PA 19096
- Purchase Order (P.O.) - for those accounts in good standing only; net 30 days
- Check/Cash - in advance of shipment
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Please have the following information ready when placing an order using a P.O., check, or cash payment:
- Contact Information (please include an email for order status notification)
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*Will be used to determine if a credit account may be opened.
Shipment is via Federal Express Standard Overnight, Monday-Thursday in the contiguous United States and Monday-Wednesday in Alaska, Hawaii, and internationally. Kits will not be shipped on a holiday or on the day prior to a holiday.
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