After addition of 100 µL D-luciferin substrate, the BLI signal from the wells was acquired in the IVIS Spectrum (Xenogen, Alameda, CA) using a highly sensitive, cooled CCD camera, and compared to the baseline signal from culture medium or whole blood or RBC lysed blood. The protein content was measured using a Pierce total protein assay (Thermo Scientific) and compared to a protein standard curve.
Pierce total protein assay
Manufactured by Thermo Fisher Scientific
The Pierce Total Protein Assay is a colorimetric assay used to determine the total protein concentration in a sample. It is based on the bicinchoninic acid (BCA) method, which involves the reduction of Cu2+ to Cu+ by proteins in an alkaline medium and the subsequent chelation of the Cu+ ions with BCA, resulting in a purple-colored complex that absorbs light at 562 nm.
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Lab products found in correlation
2 protocols using pierce total protein assay
1
Bioluminescence Imaging of Luciferase
2
Bronchoalveolar Lavage Fluid Collection
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After terminal anesthesia and exsanguination, lungs were collapsed by opening the diaphragm and tracheal insertion of 20G stub needles. A 1 mL syringe containing 1 mL of phosphate-buffered saline was then washed in and out of the lungs three times to recover bronchoalveolar lavage (BAL) fluid. BAL cells were counted using a LUNA-II automated cell counter (Logos Biosystems), and BAL supernatant total protein was measured using a Pierce total protein assay (Thermo Scientific, No. 23225).
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