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G4101-200T / 1000T MTT detection kit 1000T Cell viability detection reagent

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G4101-200T / 1000T MTT detection kit 1000T Cell viability detection reagent

This product MTT detection kit is widely used in cell proliferation and activity determination, as well as the detection of drug-to-cytotoxicity. The full name of MTT is 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide, and the Chinese name is thiazole blue. The detection principle is that the succinate dehydrogenase in the mitochondria of living cells can reduce the exogenous substrate MTT to water-insoluble blue-purple formazan (Formazan) and deposit in the cells, while dead cells have no such function. Then use dimethyl sulfoxide (DMSO) to dissolve the formazan deposits in the cells, and measure the light absorption value at 490 nm or 570 nm with a microplate reader. Within a certain range of cell numbers, the amount of formazan formation is The number of living cells is directly proportional. According to the measured absorbance value (OD value), the number of living cells can be judged. The larger the OD value, the more the number of living cells and the stronger the cell activity (if the drug toxicity is measured, it means that the drug toxicity is less). When using this kit to detect cell viability, the supernatant needs to be aspirated after the reaction, so it cannot be used for the detection of suspended cells. If you need to detect suspended cells, it is recommended to use other products of our company (G4103 CCK-8 kit).

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Product Variation:

Item code Description Volume
G4101-200T MTT detection kit 200T
G4101-1000T 1000T

 

Package Contents:

Item code Description G4101-200T G4101-1000T
G4101-1 MTT working fluid 10mL 2x25mL
G4101-2 DMSA 20mL 4x20mL
  User Manual 1pc 1pc

Technical Specification

Storage and transportation

Transport with wet ice; store at 4°C in the dark with a validity period of 3 months; or store at -20°C in the dark with a validity period of 6 months. Avoid repeated freezing and thawing.

 

Operation Steps:

  1. Adherent cell treatment: After the cells adhere to the wall, add drugs and transfection according to the experimental design;
  2. Cell viability test: add 20 μL of MTT working solution to each well and incubate for 4 h in a cell incubator;
  3. Remove the supernatant: After the incubation is completed, a small amount of purple crystals will appear at the bottom of the well plate, which can be observed under a 40x microscope. Use a pipette to carefully suck off the supernatant solution in the well (do not suck the purple crystals at the bottom);
  4. Color development: Add 100 μL DMSO to the well plate, incubate at 37°C for about 15 minutes (you can tap the bottom of the plate with your finger and shake it slightly), and wait for the purple crystals to dissolve. If the purple crystals are smaller and less, the dissolution time will be shorter; if the purple crystals are larger and more, the dissolution time can be appropriately extended;
  5. Detection: Measure the absorbance at a wavelength of 490 nm or 570 nm. If the 570 nm filter is not available, a 560-600 nm filter can be used.

 

Notes:

  1. DMSO will solidify at a lower temperature, and it can be reconstituted at 37°C without affecting its use.
  2. The detection principle of this kit relies on the reaction catalyzed by dehydrogenase. If there are more reducing agents (such as some antioxidants) in the sample that will interfere with the detection, try to remove them.
  3. For your safety and health, please wear lab coats and disposable gloves for operation.

 

The product is for scientific research purposes only, not for clinical diagnosis!

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