Protein quantification Fluorimetric method

The most widely used method for protein quantification is by spectrophotometry. The concentration of the protein in the samples is measured at an absorbance of 280 nm. The absorbance of the sample protein is then plotted against a standard curve. This method allows for total protein quantification in a sample (cell and tissue extracts). Before analysing the concentration of protein in the sample, it is important to choose the right test method.  For high protein concentration samples (above 5 - 160 mg/ml) the best method is to use the Biuret test. For low concentrations samples (between 1 - 2000µg/ml) the best methods are Lowry assay, BCA assay, Bradford assay and coomassie blue (for exact sensitivity of the test kits you use, refer to manufacturer's protocol). If the samples contain detergents like Triton X-100 then BCA assay is the best choice. For samples that have proteins larger than 3 KDa in size Bradford assay is the best choice. Each method has advantages and disadvantages, plan your analysis considering your sample characteristics.

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Found 3 matching solutions for this experiment

Protocol tips
Cells were lysed using the Bright-Glo Luciferase Assay System (Promega), and luminescence was measured on a SpectraMax L microplate reader (Molecular Devices) and normalized to protein content by NanoOrange Protein Quantification Kit (Invitrogen) using the manufacturer’s instructions with slight modifications.
EZQ™ Protein Quantitation Kit

Thermo Fisher Scientific

Protocol tips
Protein concentration was determined with the EZQ protein quantification kit (Molecular Probes, Invitrogen, Paisley, UK) or the DC Protein Assay (Bio-Rad Laboratories Sundbyberg, Sweden).
Qubit™ Protein Assay Kit

Thermo Fisher Scientific

Protocol tips
Supernatant was used for quantification of proteins and further analysis by polyacrylamide gel electrophoresis (PAGE).
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