Advanced Protein Assay (ADV01)

Material
The Advanced Protein Assay Reagent is designed to optimize the speed and accuracy of protein measurement. The reagent combines the useful properties of low protein to protein variance (see Fig. 1) and a strong signal for a sensitive assay. A simple one step procedure results in a green to blue color change which can be recognized by measuring absorbance at 570 to 615 nm within 1 min.

The Advanced Protein Assay can easily be used in either a 1 cm pathlenght cuvette format or in a 96-well format. The accompanying manual contains instructions for several variants of both measuring options as well as an extensive troubleshooting guide.

The Advanced Protein Assay Reagent is ideally suited to protein solutions in the range of 25 µg/ml to 1000 µg/ml. For more highly concentrated proteins or a wider range of protein concentrations (0.25 mg/ml to 50 mg/ml) it is recommended to use the Precision Red protein assay (Cat. # ADV02).

Figure 1: The low protein to protein variance with ADV01 and ADV02. A comparison of ADV01, ADV02, Bradford and Lowry assays on five different proteins.

Detergent and chemical compatibility
The Advanced protein Assay is a highly stable assay and it tolerates a wide range of reagents. See the Advanced Protein Assay reagent compatibility page for a full list of reagents that have been tested.

One example of specialized use for the Advanced Protein Assay is to measure protein concentrations in SDS-PAGE sample buffer, easily allowing you to control how much protein you are loading on your SDS-PAGE gels. See Figure 2 below.

Figure 2. Comparison of protein measurements with ADV01 in PBS buffer and SDS-PAGE sample buffer. 2 µl of sample plus 300 µl of reagent were mixed in a well of a standard 96-well plate. Absorbance at 600 nm was read and plotted on the figure. ADV01 gives a linear response in both PBS and SDS-PAGE sample buffer. Note: The coefficient does change and measuremements in SDS-PAGE sample buffer should be compared to standards in the same buffer.

Examples of publications where this product was used
Gong, F., Ito, K., Nakamura, Y. and Yanofsky, C. (2001). The mechanism of tryptophan induction of tryptophanase operon expression: tryptophan inhibits release factor-mediated cleavage of TnaC-peptidyl-tRNA(Pro). Proc. Natl. Acad. Sci. U. S. A. 98, 8997-9001.

Honnappa, S., Cutting, B., Jahnke, W., Seelig, J. and Steinmetz, M. O. (2003). Thermodynamics of the Op18/stathmin-tubulin interaction. J. Biol. Chem. 278, 38926-38934.

Mohri, K. and Ono, S. (2003). Actin filament disassembling activity of Caenorhabditis elegans actin-interacting protein 1 (UNC-78) is dependent on filament binding by a specific ADF/cofilin isoform. J. Cell Sci. 116, 4107-4118.

Rizzo, M. A. and Piston, D. W. (2003). Regulation of β cell glucokinase by S-nitrosylation and association with nitric oxide synthase. J. Cell Biol. 161, 243-248.

Schweizer, A., Briand, C. and Grutter, M. G. (2003). Crystal structure of caspase-2, apical initiator of the intrinsic apoptotic pathway. J. Biol. Chem. 278, 42441-42447.