Description: SRB Viability/Cytotoxity Assay Kit is a colorimetric assay based upon the quantitative staining of cellular proteins by sulforhodamine B (SRB). The SRB assay provided a better linearity with cell number and a higher sensitivity. Cell debris is not stained by SRB, therefore the sensitivity of SRB assay for cytotoxicity detection is not affected by cell debris. This assay is accurate, simple, reliable and reproducible. It has been widely used in cytotoxicity and cell viability studies. 

 Kit Contents for 1000 Assays: 

1. 1.fixation solution 60ml 

2. 1. SRB staining solution 60ml 

3. 10.dye wash solution 60ml 

4. 1.dye solubilization solution 60ml 

Storage 

Room temperature 

Material Needed But NOT Supplied with the Kit 

96 well tissue culture plates 

Microplate reader 

Preparation before Starting 

Dilute the 120ml of 10.dye wash solution by adding 1080ml distilled water 

 Protocol: 

1. Cells are grown in a 96-well plate in 200 μl growth medium. 

Note: In general, 100~ 5000 cells per well are acceptable for detection. 

2. Without removing the cell culture supernatant, gently add 50 μl cold 1.fixation solution to each well, and incubate the plates at 4 °C for 1 h. 

Note: The plates should be disturbed as little as possible during and after Fixation solution step.Do not inject the water stream directly onto the bottom of the wells, as this can cause the cell monolayer to detach. 

3. Wash five times with tap water after fixation. 

4. Cells are stained by addition of 50 μl/well of 1. SRB staining solution at room temperature for 30 min, and rinsed four times with 200 μl (each time) of 1.dye wash solution to remove unbound dye. 

Note: It is very important to rinse the plates several times, and the rinses should be carried out as quickly as possible 

5. SRB was solubilized in 50 μl of 1.dye solubilization solution for 5 min with agitation, 

6. Measure absorbance at 560~580 nm read with a microplate reader. 

Note: This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals. Avoid contact with eye,skin and clothing. Do not ingest. Wear gloves.