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Absorbance readings of samples were made at 420 nm and corrected for blank-well absorbance.
Optical density readings of samples were converted to relative levels using values obtained from standard curves generated with serial dilutions of each recombinant cytokine.
We converted optical density readings of samples to milligrams per milliliter using values obtained from a standard curve generated with serial dilutions of bovine serum albumin (0.1 1.5 mg/mL).
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Each pressure and temperature level was kept constant for 5 min to get stable readings of sample resistance.
The medium backgrounds of absorbance and fluorescence were determined from blank wells loaded with LB media and were subtracted from the readings of sample wells.
The background reading was subtracted from the readings of the samples, and PARP activity was calculated using the standard curve obtained from the readings of standards.
For more accurate measurements, the absorbance of the no-cell controls was subtracted from the readings of CM samples.
Readings of all samples were performed immediately (t = 0 min) and after 30 min or 120 min of incubation.
This information was then used to correct by subtraction the spectrophotometric readings of effluent samples in experiments with colloids.
The instrument took fluorescence readings of all samples at regular intervals of 50 min for a total duration of 92 h.
We used a standard ELISA reagent kit (OptEIA Reagent Set B; BD-Pharmingen) and converted OD450 readings of the samples to EU.
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