I. Introduction:
The VisionBlueTM Quick Cell Proliferation Assay Kit provides by far the most sensitive and easiest means for quantifying cell proliferation and viability. Simply add the single reagent to cell culture, incubate and read the fluorescence intensity. The VisionBlueTM assay utilizes the redox dye resazurin which is not fluorescent, but upon reduction by viable metabolically active cells, the dye becomes highly fluorescent (Ex = 530-570 nm; Em = 590-620 nm). Therefore, viable metabolically active cells can be easily measured.
Key Features:
1. Simple procedure: Just Add-Incubate-Read. No washing, No solublization.
2. Highly Sensitive: Detect as few as 100 cells.
3. Large linear assay range: Detect from 100 to 100000 cells per well.
4. Longer Stability: Reagent is stable at 4o C for at least a year.
5. Very safe: Non-radioactive, non-toxic, and cells can be further used for other experiments.
6. Wide applications: Used in studying a variety of growth stimulations or inhibitions (e.g., by growth factors, cytokine, nutrients, anticancer drugs, apoptosis inducers/inhibitors, toxicityinducing chemicals, etc.).
II. Kit Contents:
Components | K303-500 | K303-2500 | K303-Bulk |
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500 Assay | 2500 Assay | Bulk |
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VisionBlueTM Reagent | 5 ml | 25 ml | Bulk |
III. General considerations: 1. Phenol red or serum does not interfere with the VisionBlueTM assay.
2. The assay can be performed in any type of culture plates, adjust the VisionBlueTM reagent amount to 10% of culture medium. Duplicate or triplicate assays are recommended.
3. Drugs or compounds should be dissolved in PBS or culture medium, or perform proper solvent control if compound is dissolved in other solvents.
IV. Reagent Storage and handling:
Store the reagent at 4oC, stable for 1 year, protect from light. For research use only! V. Cell Proliferation Assay Procedures: 1. Plate cells (1-50x104/well) in a 96-well microtiter plate in a final volume of 100 µl/well culture medium. For toxicity assays, use more cells to start with (e.g., 1-5 x 105 cells/well). Notes:
(1) The optimal cell number used for the assay may vary among cell types. For best results, it is recommended to add various numbers of cells in your initial assay to determine the optimal cell number to be used.
(2) We recommend performing a reagent fluorescence background control by using the same amount of culture medium and VisionBlueTM Reagent without any cells.2. Treat cells with your stimuli or drug for desired period of time (e.g. 12-96 hours).
3. Accurately add 10 µl (10% medium volume) VisionBlueTM Reagent into each well, mix well gently. Be careful not to introduce bubbles to the wells.
4. Incubate the plate for 1 - 5 hours in standard culture conditions.
Note: Incubation time is dependent on cell type and cell number used. You may read the plate multiple times as desired and choose the best reading results.
5. Measure fluorescence intensity on a fluorescence plate reader, or fluorometer at Ex. = 530-570 nm, Em. = 590-620 nm.
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