Sulforhodamine B Assay: A Sensitive Assay to Measure Drug Resistance Via Determination of Cellular Protein Content in Cancer Cells
Published: April 30, 2023
Abstract
Source: Sciarrillo, R. et al. Using RNA-sequencing to Detect Novel Splice Variants Related to Drug Resistance in In Vitro Cancer Models. J. Vis. Exp. (2016)
This video describes the high throughput colorimetric screening assay, sulforhodamine B assay, to measure drug-induced cytotoxicity in cancer cells. The assay determines the drug resistance by measuring the cell density based on the cellular protein content.
Protocol
1. SRB Assay for Pancreatic Carcinoma Cells Dissolve SRB reagent in 1% acetic acid at final concentration of 0.4% (w/v). Dissolve trichloroacetic acid (TCA) in ultrapure water at a final concentration of 50% (w/v). Dissolve Tris(hydroxymethyl)-aminomethane in ultrapure water at a final concentration of 10 mM. Prepare a separate 96-well flat-bottom "Day 0" control plate to ensure more accurate estimation of growth inhibition: Seed 6 wells with cells growin…
Divulgaciones
The authors have nothing to disclose.
Materials
| Sulforhodamine B | Sigma-Aldrich | 230162 | |
| Trichloroacetic acid | Sigma-Aldrich | 251399 | |
| Panc-1 | ATCC, Manassas, VA, USA | ATCC CRL-1469 | |
| Tris(hydroxymethyl)-aminomethane | Sigma Aldrich | 252859 | |
| Greiner CELLSTAR 96 well plates | Greiner/Sigma | M0812-100EA | |
| Anthos-Elisa-reader 2001 | Labtec, Heerhugowaard, Netherlands | UV-Vis 96-well plate spectrophotometer | |
| RPMI-1640 | Gibco, Carlsbad, CA, USA | 11875093 | |
| Greiner CELLSTAR 96 well plates | Greiner/Sigma | M0812-100EA | |
| Fetal bovine and calf serum | Greiner Bio-One, Frickenhausen, Germany | 758093 | |
| penicillin G streptomycin sulphate | Gibco, Carlsbad, CA, USA | 15140122 |
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Sulforhodamine B Assay: A Sensitive Assay to Measure Drug Resistance Via Determination of Cellular Protein Content in Cancer Cells. J. Vis. Exp. (Pending Publication), e20492, doi: (2023).