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Quantification made easy

io A260/A230 should be approximately 2.2. 2. Protein determination

The protein content of a preparation can be determined on the basis of various different analytical procedures. Evaluation can be carried out via factor or via a calibration curve, with up to ten standards in the BioPhotometer. Absorption measurement at 280 nm (A280)
A280 method may be used with in concentrations of up to approximately 4 mg/ml (3.0 A). This method is simple and rapid, but may be disturbed by the parallel absorption of non-proteins (e.g. DNA). Unlike the colorimetric process, this method is less sensitive and requires higher protein concentrations and should thus be used with pure protein solutions. In addition to the direct absorbance display, evaluation is possible with the BioPhotometer via the Warburg formula or via standard. Colorimetric determination (dye tests)
Protein samples often consist of a complex mixture of many different proteins. The quantitative detection of the protein content is usually achieved on the basis of the reactions shown by functional groups of the proteins to dye-forming reagents. The intensity of the dye correlates directly with the concentration of the reacting groups and can be measured exactly. Lowry assay 595nm
Specialist literature contains a multitude of modifications for the Lowry assay. The principal target is to reduce the high susceptibility to interference. In comparison to the pure Biuret assay, the sensitivity of this assay has greatly increased. However, the Lowry method is adversely affected by a wide range of
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