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Star Republic: Guide for Biologists

Measure cell proliferation --- sulforhodamine B

Virtualab Protocols (Chang Bioscience)
1. Seed 100 l (5000 - 40,000 cells) per well in 96 well plate.

2. You may need to add dilution of drugs or other agents at this time or after cells have attached overnight.

3. Incubate cells for the desired time at 37 C.

4. To fix cells, add 50 ml of cold 50% TCA (adherent cells) or 80% TCA (suspension cells) to each well dropwise.

5. Fix at 4 C for 1-3 hours.

6. Remove the liquid by inverting the plate.

7. Rinse the plates 5 times with water, tapping on paper towels after rinsing.

8. Air dry the plates (face up) in room temperature for 12-24 hours.

9. Add 100 l 0.4% sulforhodamine B (SRB)/1% glacial cetic acid to each well.

10. Incubate at room temperature for at least 5 minutes.

11. Rinse the plates 3 times with 1% glacial acetic acid, tapping on paper towels after rinsing.

12. Air dry the plates (face up) in room temperature for 12-24 hours.

13. Add 100 l of 10 mM Tris and shake until all bound SRB is into the solution.

14. Read on a microplate reader at 515 nm.