MTT cell proliferation and cytotoxicity test kit
MTT cell proliferation and cytotoxicity test kit
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| Usage | 1. Place the cell plate in the sterile operating table. 2. Add 10% volume of MTT solvent to each well of the cell plate. [Preparation of MTT solution: Dissolve 25mg of MTT with 5ml of MTT solvent to prepare a 5mg/ml MTT solution. It can be used after preparation, or stored directly at-20 ℃ in the dark from light, or stored at-20 ℃ in the dark from light after appropriately packaging as needed.] 3. Put the cell plate back into the incubator and continue to incubate for 3-4 hours. When doing comparative experiments, the incubation time should be consistent. 4. After incubation, take out the cell plate from the incubator and dissolve it according to the following method to produce MTT formazan crystals. (1) If it is an adherent cell, discard the culture medium. Add the same amount of Formazan Solvent as the initial volume. The volume of dissolved fluid may vary per well, but the total volume must be consistent for comparison. (2) If the removal of non-adherent cells or culture medium will cause the loss of MTT formazan, add the same amount of lysate as the initial volume directly to the culture medium. (3) Readings must be completed within 1 hour after addition of Formazan Solvent. 5. Gentle shaking in a shaker can enhance dissolution. In some cases, particularly when the cell density is high, repeated pipetting is required to completely dissolve the MTT formazan crystals. 6. Spectrophotometer to measure OD570 and reference wavelength OD690. 7. The 96-well plate can be detected on a microplate reader equipped with a suitable filter. 8. Other types of multi-well plates need to be spectrophotometrically detected with a cuvette or plate reader of appropriate size. |
| Beilstein | 0 |
| Synonym | MTT Cell Proliferation and Cytotoxicity Assay Kit,MTT kit |
| Description |
The MTT assay, also known as MTT colorimetry, is a method to detect cell survival and growth. The detection principle is that succinate dehydrogenase in mitochondria of living cells can reduce exogenous MTT to water-insoluble blue-violet crystalline Formazan and deposit it in cells, while dead cells do not have this function. Within a certain range of cell numbers, the amount of MTT crystals formed is directly proportional to the number of cells. The greater the absorbance, the stronger the cell activity, indicating that the drug toxicity is less. This method has been widely used in the activity detection of some bioactive factors, large-scale anti-tumor drug screening, cytotoxicity test and tumor radiosensitivity determination. It is characterized by high sensitivity and economy. Components: MTT (25mg), MTT solvent (5ml), Formazan solution (55ml)
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| PubChem CID | 0 |
| General Notes | 1. Since a 96-well plate is used for detection, if the cell culture time is long, we must pay attention to the problem of evaporation. On the one hand, since the circle around the 96-well plate is the easiest to evaporate, you can discard the circle around the plate and add PBS, water or culture medium instead; On the other hand, a 96-well plate can be placed close to a water source in the incubator to alleviate evaporation. 2. If the test compound used can react with MTT, centrifuge first and then discard the culture medium. Carefully flush it with PBS for 2 to 3 times, and then add the culture medium containing MTT. 3. The production of Formazan is not only proportional to the number of living cells, but also affected by the action time. Therefore, when there are many samples, the measurement results may change with time. 4. This product contains carcinogens, avoid contact with skin and eyes, please operate carefully. For your safety and health, please operate in a lab coat and disposable gloves. 5. This product is only used for scientific research and cannot be used clinically or diagnostically. |
| Storage Temp. | Stored at 2 ~ 8 ℃, protected from light, valid for 12 months. |
