Protein Assay Kit, BioAssay(TM)

The protein is known as the "building blocks of life" and is one of the most important macromolecules in life science. Proteins are polypeptides made up of amino acids and play various key roles in all aspects of biology. Protein quantitation is a very common practice for life scientists.\n\nSimple, direct and automation-ready procedures for measuring protein concentration are very desirable. Protein assay kit is based on an improved Coomassie Blue G method. The dye forms a blue complex specifically with protein, and the intensity of color, measured at 595nm, is directly proportional to the protein concentration in the sample. The optimized formulation substantially reduces interference by substances in the raw samples and exhibits increased sensitivity towards peptides.\n\nApplications:\nDirect Assays: total protein concentration.\n\nKey Features:\nSensitive and accurate. Use 10ul samples. Detection range 0.06пїЅ1.0mg /mL protein in 96-well plate assay.\nSimple and high-throughput. The пїЅmix-and-readпїЅ procedure involves addition of a single working reagent and reading the optical density. Can be readily automated as a high-throughput assay in 96-well plates for thousands of samples per day.\nLow interference. Glucose, Tris, vitamins, and amino acids, DNA, RNA, salts, EDTA (< 12mM), phenol (< 50mM), urea (< 0.6 M), Triton (< 0.1%) and SDS (< 0.1% SDS) do not interfere in the assay.\nVersatility: assays can be executed in 96-well plate or cuvet.\n\nKit Contents: (500 tests in 96-well plates)\nReagent: 20ml 5 x concentrate\nProtein standard: 1ml 1.0 mg/mL BSA\nStorage conditions. The kit is shipped at room temperature. Store the reagent at 4 degrees C and standard at -20 degrees C, respectively. Shelf life: 12 months after receipt.\nPrecautions: reagents are for research use only. Normal precautions for laboratory reagents should be exercised while using the reagents. Please refer to Material Safety Data Sheet for detailed information.\n\nProcedures:\nReagent Preparation:\nPrepare enough working reagent by adding 1 vol of the 5 x Reagent to 5 vol of distilled water. Bring reagent to room temperature before use.\nProcedure using 96-well plate:\n1. Dilute standard as shown in the Table. Transfer 10ul diluted Standards and diluted sample in duplicate wells of a clear bottom 96-well plate. Store diluted standards at -20 degrees C for future use.\nNo Premix+H2O Vol (ul) BSA (mg/mL) BSA (ug/10ul)\n1 100ul+0ul 100 1.0 10\n2 80ul+20ul 100 0.8 8\n3 60ul+40ul 100 0.6 6\n4 40ul+60ul 100 0.4 4\n5 30ul+70ul 100 0.3 3\n6 20ul+80ul 100 0.2 2\n7 10ul+90ul 100 0.1 1\n8 0ul+100ul 100 0 0\n2. Add 200ul working reagent and tap lightly to mix.\n3. Measure OD at 570-630nm (peak 595nm).\nProcedure using cuvette:\n1. Prepare standards as in the 96-well plate assay. Transfer 50ul diluted Standards and 50ul samples to cuvets.\n2. Add 1000ul working reagent and tap lightly to mix.\n3. Measure OD at 570-630nm (peak 595nm).\n\nCalculation:\nSubtract blank OD (water, #8) from the standard OD values and plot the OD against standard concentrations. Use the standard curve to determine the sample protein concentration.\n\nMaterials Required, But Not Provided:\nPipeting devices and accessories.\nProcedure using 96-well plate:\nBlank 96-well plates (e.g. Corning Costar).\nPlate reader for 96-well plate.\nProcedure using cuvette:\nCuvets and spectrophotometer.\n\nGeneral Considerations:\nIf protein concentration is > 1 mg/mL, dilute samples in distilled water, and use OD values that lie within the calibration curve to calculate the sample protein concentration. Reading can be performed as soon as the reagent and sample are mixed. High sensitivity can be achieved by adding 50ul sample to 200ul Reagent (detection range 3пїЅ200 ug/mL).

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SPECIFICATIONS

Catalog Number

P9102-20P

Size

1Kit

References

1. Sharifuzzaman, S.M. and Austin, B. (2009). Kocuria SM1 controls vibriosis in rainbow trout (Oncorhynchus mykiss, Walbaum). J Appl Microbiol. 108(6): 2162-2170.\n2. Stefanidis A., et al. (2009). The role of thermogenesis in antipsychotic drug-induced weight gain. Obesity (Silver Spring). 17(1): 16-24.\n3. Verty, A.N. et al. (2009). The effects of rimonabant on brown adipose tissue in rat: implications for energy expenditure. Obesity (Silver Spring). 17(2): 254-61.