Boster Immunoleader

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Neutral Red Cell Proliferation and Cytotoxicity Assay Kit

AR1157

500T

50 USD

AR1157

Neutral Red Cell Proliferation and Cytotoxicity Assay Kit

500T

50 USD

Assistant Reagent

At -20˚C for one year. Neutral red staining solution should be stored in dark.

Neutral Red Cell Proliferation and Cytotoxicology Assay Kit base on the cells’ ability of absorbing neutral red to detect cell proliferation an cytotoxicology. Neutral red can be absorded by alive cells and accumulate in lysome. When cell proliferation speed up, as the cells become more, the quantity of neutral red are absorbed more; once cell was damaged, the ability of absorbing neutral red may disappear.In this way, we can judge the situation of cell proliferation an cytotoxicology. Meanwhile, neutral red also is a PH indicator. It is red in acid, when PH from 6.8 raise to 8.0, it will turn to yellow. Because nucleus is acid, it can be stained to yellow. And the circumstance in lysome is acid either; it can bestained to red by neutral red. The molecular formula of neutral red is C15H17IN4 with 288.8 KDa molecular weight. This Kit is for 500 times experiment (five 96-wells).

1. Culture cells in 96 wells, add 200ul cell culture fluid into each well, then stimulate with drug. 2. Add 20ul neutral red solution if the drug in cell culture fluid will not interrupt the detection; if it will do, please wash sample 2-3 times with PBS, DPBS or HBSS buffer, then add 200ul cell culture fluid and 20ul neutral red solution. 3. Incubate for 2 hours.（if the cell density is very law and the cell metabolic rate is very slow, you may last 3-4 hours）. 4. Wash sample with PBS, DPBS or HBSS buffer 1-2 times to remove the cell culture fluid. 5. Add 200ul neutral red detecting solution, disintegrate on shaker for 10 min at room temperature. 6. Measure sample’s A450. Suggest choose 690nm as wavelength. 7. Set blank wells and contrast wells. Note 1. Prepare an ELISA or a micro-spectrophotometer that can detect A540. 2. Do preliminary experiment before first experiment. 3. Please wear transparent gloves when operate. Tips 1. After a long time storage, neutral red solution may appear some sediment. Use the supermatant or remove sediment after filter. It won’t interrupt the result. 2. When cell culturing, 96 wells may appear evaporate issue. It will interrupt cells growth situation and interrupt the result badly.

For quantitation of viable cell number in proliferation and cytotoxicity assays

200ul per time, and can be used about 500 times