Catalog No.: CSK1157Kit ComponentNeutral red staining solution 10mlNeutral red detective solution 100mlStorageAt -20℃ for one year.Neutral red staining solution should be stored indark.IntroductionNeutral Red Cell Proliferation and CytotoxicologyAssay Kit base on the cells’ ability of absorbingneutral red to detect cell proliferation ancytotoxicology. Neutral red can be absorded byalive cells and accumulate in lysome. When cellproliferation speed up, as the cells become more,the quantity of neutral red are absorbed more;once cell was damaged, the ability of absorbingneutral red may disappear.In this way, we canjudge the situation of cell proliferation ancytotoxicology. Meanwhile, neutral red also is aPH indicator. It is red in acid, when PH from 6.8raise to 8.0, it will turn to yellow. Because nucleusis acid, it can be stained to yellow. And thecircumstance in lysome is acid either; it can bestained to red by neutral red.The molecular formula of neutral red isC15H17IN4 with 288.8 KDa molecular weight.This Kit is for 500 times experiment (five 96-wells).
Protocol1. Culture cells in 96 wells, add 200μl cell culture fluid into each well, then stimulate with drug.2. Add 20μl 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 200μl cell culture fluid and20μl neutral red solution.3. Incubate for 2 hours. if the cell density is very law and the cell metabolic rate is very slow, you maylast 3-4 hours.4. Wash sample with PBS, DPBS or HBSS buffer 1-2 times to remove the cell culture fluid.5. Add 200μl 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.Note1. 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.Tips1. After a long time storage, neutral red solution may appear some sediment. Use the supermatant orremove 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 andinterrupt the result badly.