MTT Assay Kit

Product Information

Product Description/Introduction

This product MTT detection kit is widely used in the determination of cell proliferation and activity, as well as the detection of drug 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 thiazolyl blue. The principle of the assay is that the enzyme succinate dehydrogenase in the mitochondria of living cells can reduce the exogenous substrate MTT to water-insoluble blue-violet filth (Formazan) and deposit it in the cells, while dead cells do not have this function. Dimethyl sulfoxide (DMSO) was then used to dissolve the dirty deposits in the cells, and the absorbance values were measured at 490 nm or 570 nm using an enzyme-labeled instrument. Within a certain range of cells, the amount of formazan formed is proportional to the number of living cells. The number of living cells is measured by the absorbance value (OD value). The greater of the OD value is, the more of living cells is, and the more active of the cell is (if measured for drug toxicity, the less toxic of the drug is). If using this kit to detect cell viability, the supernatant needs to be removed after the reaction, so it cannot be used for the detection of suspended cells. For detection of suspended cells, it is recommended to use products (G4103 CCK-8 kit) in our company.

Storage and Shipping Conditions

Ship with wet ice; store at 4°C away from light, valid for 3 months; or store at -20°C away from light, valid for 12 months. Avoid repeated freezing and thawing.

Product Contents

Assay Protocol / Procedures

1. After the cells adhere to the wall, dosing and transfection are carried out according to the experimental design;

2. Add 20 μL of MTT working solution to each well and incubate for 4 h in a cell incubator.

3. Carefully remove the supernatant, do not suck up the purple crystals at the bottom.

Note: After incubation, a small amount of purple crystals will appear at the bottom of the well plate, which can be observed under a 40x microscope.

4. Add 100 μL of DMSO to the well plate, incubate at 37°C for about 15 minutes (you can tap the bottom of the plate with your finger to shake slightly), until all the purple crystals are dissolved. If the purple crystals are smaller and less, the dissolution time will be shorter; if the purple crystals are larger, the dissolution time can be appropriately extended;

5. Read the absorbance at 490 nm or 570 nm. If a 570 nm filter is not available, a 560-600 nm filter can be used.

Note

1. Prior to use, precipitates that might be formed at lower temperature in DMSO should be resolubilized completely at 37 in a water bath.

2. This kit employs the reduction reaction catalyzed by dehydrogenase. Reducing agents or antioxidants that may interfere with the assay should be removed from samples to be tested.

3. For your safty and health, please wear safety glasses, gloves, or protective clothing.

For Research Use Only!

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