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Sulforhodamine B assay

AK Alexei F. Kirkin
KD Karine N. Dzhandzhugazyan
PG Per Guldberg
JF Johnny Jon Fang
RA Rikke S. Andersen
CD Christina Dahl
JM Jann Mortensen
TL Tim Lundby
AW Aase Wagner
IL Ian Law
HB Helle Broholm
LM Line Madsen
CL Christer Lundell-Ek
MG Morten F. Gjerstorff
HD Henrik J. Ditzel
MJ Martin R. Jensen
WF Walter Fischer
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Cell monolayers were incubated with cytotoxic lymphocytes for 24 h. The lymphocytes were then removed from the wells by gravity sedimentation. Briefly, the wells were filled with DPBS, sealed and turned upside-down for 20 min, after which the sealer was detached without changing the plate’s position, and excess liquid removed by gentle blotting on a paper towel. Adherent cells were fixed, stained with 0.06% Sulforhodamine B (SRB) and processed according to a previously described version45 of the SRB assay46. The dye was extracted and absorbance measured at 550 nm for cultures incubated with or without lymphocytes. The survival index was determined as the ratio between these cultures, with the control culture (without lymphocytes) set as 100%.

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