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).

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