SRB Viability/Cytotoxicity 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 kit is designed for 4000 SRB Viability/Cytotoxicity Assays.This assay is accurate, simple, reliable and reproducible. It has been widely used in cytotoxicity and cell viability studies.
- VB-4000L-1 1×fixation solution———————–240ml
- VB-4000L-2 1× SRB staining solution—————240ml
- VB-4000L-3 10×dye wash solution——————240ml
- VB-4000L-4 1×dye solubilization solution——–240ml
Store at 2-8ºC.
Material Needed But NOT Supplied with the Kit
- 96 well tissue culture plates
- Microplate reader
Preparation before Starting
Dilute the 100ml of 10×dye wash solution by adding 900 ml distilled water
- 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.
- 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.
- Wash five times with tap water after fixation.
- 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.
- SRB was solubilized in 50 μl of 1×dye solubilization solution for 5 min with agitation.
- 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 eyes, skin and clothing. Do not ingest. Wear gloves.