I am working on melanoma B16-F10 cell line and after treatment whit different concentration of drug I do SRB protocol then I read their absorb at wavelengths of 545 nm. now, how to analyze these results? What should be the range of control absorbency?
Hi
You can use the following protocol to calculate IC 50 for your drug. PDF attached for your reference.
The SRB assay is an arbitrary unit comparative technique. That is, the end result of your treatments should be compared to your negative control. The average of the controls is considered as 100% of the cell viability. For example, say we have the average of the controls being 50, you will divide each sample value by 50 and multiply by 100. After that, all data will be in percentage compared to the control.
[Sample Value/Controls Mean X 100]
Remember to substract the brackground of each sample (for example, empty wells). Since you are working with multiple doses, it is interesting to calculate the IC50 value. To learn more about, I recommend this introductory article;
https://www.graphpad.com/support/faq/50-of-what-how-exactly-are-ic50-and-ec50-defined/
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