Principle

Fast, sensitive, and convenient assay for protein quantification

The Protein Quantification Assay is a convenient and reliable kit for the determination of protein concentration in buffers typically used for SDS-PAGE (sodium dodecylsulfate polyacrylamide gel electrophoresis). It is mainly designed for proteins solved in Protein Solving Buffer (PSB, a component of NucleoSpin® RNA/Protein, formerly named PLB) or in buffer as described by Laemmli (1970). These buffers usually contain SDS, a reducing agent, a dye, and a component to increase the buffer density. The majority of protein quantification assays are either influenced by or incompatible with SDS, reducing agents, or dyes, commonly present in protein sample buffers. The MACHEREY-NAGEL Protein Quantification Assay however, is well suited for these buffer systems. It is a fast and sensitive assay, based on a modification of a protocol described by Karlsson et al. (1994). Protein quantification can be carried out in microplates, microcuvettes, semi-microcuvettes, and with low-volume photometers (e.g., NanoDrop™).

Features

Procedure

1. BSA dilution series
   
   
2. Protein samples
   
   
3. Addition of Quantification Reagent QR to the dilution series and to the samples.
   Turbidity appears corresponding to the protein concentration.
   
   
   
   
4. Measuring of the turbidity of the dilution series by light extinction measurement at 570 nm to create a calibration curve (concentration vs. extinction).
5. Measuring of the turbidity of the protein samples by light extinction measurement at 570 nm for determining protein concentration by comparing with the calibration curve.