Neutral red Cell Proliferation and Cytotoxicology Assay Kit

Description

Catalog No.: CSK1157
Kit Component
Neutral red staining solution 10ml
Neutral red detective solution 100ml
Storage
At -20℃ for one year.
Neutral red staining solution should be stored in
dark.
Introduction
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 be
stained 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).

Protocol
1. 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 and
20μl 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 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.
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.