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Index / Bioassays / CellQuanti_Blue™ Cell Viability Assay Kits / Product Detail : CQBL-05K CellQuanti_Blue™ Cell Viability Assay Kits
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#CQBL-05K CellQuanti_Blue™ Cell Viability Assay Kits

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  Price : 239   EUR
272   USD
185   GBP
1006   Zloty
32063   JPY
1848   NOK
1980   SEK
271   CHF

Product name : CellQuanti_Blue™ Cell Viability Assay Kits

Catalog number : CQBL-05K

Quantity: 5000

Availability: Yes

Supplier name : Bioassays

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About this Product :

CellQuanti_Blue™ Cell Viability Assay Kits antibody storage GENTAUR recommends for long therm storage to freeze at -24 C. For short time storage up to 30 days we suggest fridge storage at 1 to 10 C. Prevent multiple freeze taw cycles of CellQuanti_Blue™ Cell Viability Assay Kits.
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More Details about

This homogeneous assay involves simply adding a single reagent, the CellQuanti-BlueTM reagent, to the cell culture and measuring the fluorescence intensity (excitation wavelength = 530 - 570 nm, emission wavelength = 590 - 620 nm) after an incubation step. The CellQuanti-BlueTM reagent, like other resazurin-based assays such as the Alamar Blue reagent, utilizes the redox dye resazurin which is not fluorescent, but upon reduction by metabolically active cells is converted into a highly fluorescent
product (resorufin). Living cells can readily reduce this non-toxic reagent and the resulting increase in fluorescence intensity can be conveniently monitored using a fluorescence spectrophotometer or plate reader.


Nonviable cells have no metabolic capacity and, thus, will not reduce the dye. Therefore, the fluorescence intensity observed in this assay is a true measure of the viable cells. The CellQuanti-BlueTM reagent has been optimized for maximum sensitivity, reproducibility and long shelf-life. The homogeneous cell-based assay can be performed in multi-well plates. The reagent is compatible with all culture media and with all liquid handling systems for high-throughput screening applications in 96-well and 384-well plates. Applications include cell proliferation, cytotoxicity and apoptosis.

Safe. Non-radioactive assay (cf. 3H-thymidine incorporation assay).
Sensitive and accurate. As low as 100 cells can be accurately quantified.
Time efficient. High-throughput assay in 96-well and 384-well plates allows simultaneous processing ten of thousands of samples per day.
Homogeneous and convenient. A single reagent and "mix-incubatemeasure" type assay. No wash and reagent transfer steps are involved.
Robust and amenable to HTS: Z’ factors of 0.6 to 0.9 are routinely observed in 96-well and 384-well plates. Can be readily automated on HTS liquid handling systems.

Cell Proliferation: effects of cytokines, growth factor, nutrients.
Cytotoxicity and Apoptosis: evaluation of toxic compounds, anti-cancer antibodies, toxins, environmental pollutants etc.
Drug Discovery: high-throughput screening for anticancer drugs.

Storage conditions. The CellQuanti-BlueTM reagent is light sensitive and should be stored in the provided amber bottle at 4°C. The Control Reagent is stable at -20°C. Shelf life: 12 months after receipt.

Precautions: reagents are for research use only. Normal precautions for laboratory reagents should be exercised while using the reagents. Please refer to Material Safety Data Sheet for detailed information.

PROCEDURES
The CellQuanti-BlueTM assay is based on the conversion of the nonfluorescent reagent to fluorescent product by metabolically active cells. For most cells this reductive reaction takes 1 to 5 hours. The fluorescence intensity of the product is then quantified on a fluorescent microplate reader. Although most culture media contain phenol red, phenol red does not interfere with the assay. All data in Technical Notes were obtained in culture media containing phenol red.


Reagent Preparation:
Important: bring reagent to room temperature before use.

Procedure using 96-well plate:
1. Plate and culture cells (80 μL) in black 96-well tissue culture plates. Typical culture medium contains DMEM, 10% fetal bovine serum and antibiotics (penicillin/ strepto-mycin, gentamycin, etc), amino acids and other nutrients. Assays can be performed on either adherent cells or cells in suspension. The number of cells can vary from 100 to 80,000 per well. The volume can vary from 50 to 150 μL, although 80 μL is used in this protocol. In addition to the test samples, control wells of culture medium containing no cells or cells treated with a toxic reagent such as 0.1% saponin should be included.
2. Add test compounds and controls and incubate cells for the desired period of time (typically overnight). It is recommended that assays be run in duplicate or triplicate. Compounds and controls (20 μL) can be added in phosphate buffered saline (PBS) or culture medium. The Control reagent can be conveniently reconstituted with 5 mL PBS (1% saponin).
3. Equilibrate the CellQuanti-BlueTM Reagent to room temperature. Add 10 μL (per 100 μL of cell culture) of the reagent per well. The volume of the reagent can be adjusted depending on the volume of cell culture. Tap plate to mix cells with compounds. Incubate for 1 to 5 hours at 37°C.
4. Measure fluorescent intensity for each well on a fluorescence plate reader. If a Molecular Devices LJL Analyst is used, use the rhodamine filter sets (530nm excitation filter, 590nm emission filter and 570nm
dichroic mirror).

Procedure using 384-well plate:
1. Plate and culture cells (40 μL) in black 384-well tissue culture plates.
The number of cells can vary from 100 to 20,000 per well. The volume can vary from 25 to 60 μL, although 40 μL is used in this protocol. In addition to the test samples, control wells of culture medium containing no cells or cells treated with a toxic reagent such as 0.1% saponin should be included.
2. Add test compounds and controls and incubate cells for the desired period of time. It is recommended that assays be run in duplicate or triplicate and that compounds be added in PBS or culture medium with a volume of 10 μL.
3. Equilibrate CellQuanti-BlueTM Reagent to room temperature. Add 5 μL Reagent (per 50 μL of cell culture). Tap plate lightly to mix Reagent with cells. Incubate for 1 to 5 hours at 37°C.
4. Measure fluorescent intensity for each well on a fluorescence plate reader. If a Molecular Devices LJL Analyst is used, use the rhodamine filter sets (530nm excitation filter, 590nm emission filter and 570nm
dichroic mirror).

 

GENERAL CONSIDERATIONS
Incubation time. The incubation time is dependent on the cell line. Some cell lines exhibit strong metabolic activity and, thus, require shorter incubation time than less metabolically active cell lines. The incubation time can be easily determined by reading the plate multiple times e.g. every 30 minutes after adding the CellQuanti-BlueTM reagent. In general, incubation for 1 to 5 hours is sufficient. Extensive incubation (such as >18 hours) may result in non-linear fluorescence response at high cell numbers.
Cell number. Generally the optimized CellQuanti-BlueTM reagent shows a broad range linear fluorescence response to the number of culturing cells. It is recommended to determine the number of cells per well that gives a highest signal:noise ratio. The optimal cell number can be easily determined by serial dilution of cells.
Controls. A positive control that is either cytotoxic or promotes cell proliferation can be run although it is not required. Saponin is a cytotoxic detergent that is available from BioAssay Systems (see Figure 2 in
Technical Notes). A blank control, i.e., culture medium without cells or cells containing 0.1% saponin, should be done for each assay. The blank control determines background fluorescence that must be subtracted for data analysis.

DATA ANALYSIS
For cell proliferation or cytotoxicity assays, the activity of a test compound can be calculated as percent change in cell number as follows, Activity (%) or Cell viability (%) = 100 x (Fcmpd - Fo) / (Fctrl - Fo) where Fcmpd and Fctrl are the average fluorescence intensities in the presence and absence (vehicle control) of the test compound and Fo the averaged blank control fluorescence intensity.
For dose-response studies, the data are plotted against compound concentration and the EC50 for proliferation assay and IC50 for cytotoxicity assay can be determined by non-linear regression analysis using Prism or other data analysis tools.

 

TECHNICAL NOTES
The CellQuanti-BlueTM assay kit has been specially optimized and formulated to provide a sensitive, convenient and robust assay for cell proliferation and cytotoxicity. Key features of the kits are as follows:
Safe. Non-radioactive assay (cf. 3H thymidine incorporation assay).
Sensitive and accurate. As low as 100 cells can be accurately quantified.
Saves time. High-throughput assay in 96-well and 384-well plates allows simultaneous processing of a large of number of samples.
Homogeneous and convenient. A single reagent and "mix-incubatemeasure" type assay. No wash and reagent transfer steps are involved.
Robust and amenable to HTS. Z’ factors of 0.6 to 0.9 are routinely observed in 96-well and 384-well plates. Can be readily automated on HTS liquid handling systems.

 

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