Protein Assay Kit

Catalog No. ABIN1000338

Product Details

Application: Biochemical Assay (BCA)

Specificity: 60 μg/mL

Characteristics: Sensitive and accurate. Use 10 µL samples. Detection range 0.06 - 1.0 mg /mL protein in 96-well plate assay.
Simple and high-throughput. The mix-and-read 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.
Low interference. Glucose, Tris, vitamins, and amino acids, DNA, RNA, salts, EDTA (< 12 mM), phenol (< 50 mM), urea (< 0.6 M), Triton (< 0.1%) and SDS (< 0.1% SDS) do not interfere in the assay.
Versatility: assays can be executed in 96-well plate or cuvet.

Components: Reagent: 20 mL 5 x concentrate. Protein standard: 1 mL 1.0 mg/mL BSA.

Material not included: Pipeting devices and accessories. Blank 96-well plates (e.g. Corning Costar). Plate reader for 96-well plate. Cuvets and spectrophotometer.

Application Details

Application Notes: Direct Assays: total protein concentration.

Comment:

If 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 50 μL sample to 200 μL Reagent (detection range 3 - 200 μg/mL).

Protocol: Procedure using 96-well plate:
1. Dilute standard. Transfer 10 µL diluted Standards and diluted sample in duplicate wells of a clear bottom 96- well plate. Store diluted standards at -20°C for future use.
2. Add 200 µL working reagent and tap lightly to mix.
3. Measure OD at 570-630nm (peak 595nm).

Procedure using cuvette:
1. Prepare standards as in the 96-well plate assay. Transfer 50 µL diluted Standards and 50 µL samples to cuvets.
2. Add 1000 µL working reagent and tap lightly to mix.
3. Measure OD at 570-630nm (peak 595nm).

Reagent Preparation:

Prepare 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.

Calculation of Results:

Subtract 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.

Restrictions: For Research Use only

Handling

Storage: 4 °C

References

Sharifuzzaman, Abbass, Tinsley, Austin: "Subcellular components of probiotics Kocuria SM1 and Rhodococcus SM2 induce protective immunity in rainbow trout (Oncorhynchus mykiss, Walbaum) against Vibrio anguillarum." in: Fish & shellfish immunology, Vol. 30, Issue 1, pp. 347-53, (2011) (PubMed).

Waheed, Al-Eknah, El-Bahr: "Some biochemical characteristics and preservation of epididymal camel spermatozoa (Camelus dromedarius)." in: Theriogenology, Vol. 76, Issue 6, pp. 1126-33, (2011) (PubMed).

Sharifuzzaman, Austin: "Kocuria SM1 controls vibriosis in rainbow trout (Oncorhynchus mykiss, Walbaum)." in: Journal of applied microbiology, Vol. 108, Issue 6, pp. 2162-70, (2010) (PubMed).

Verty, Allen, Oldfield: "The effects of rimonabant on brown adipose tissue in rat: implications for energy expenditure." in: Obesity (Silver Spring, Md.), Vol. 17, Issue 2, pp. 254-61, (2009) (PubMed).

Aldag, Gromov, García-Rubio, von Koenig, Schlichting, Jaun, Hilvert: "Probing the role of the proximal heme ligand in cytochrome P450cam by recombinant incorporation of selenocysteine." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 106, Issue 14, pp. 5481-6, (2009) (PubMed).

Sharifuzzaman, Austin: "Influence of probiotic feeding duration on disease resistance and immune parameters in rainbow trout." in: Fish & shellfish immunology, Vol. 27, Issue 3, pp. 440-5, (2009) (PubMed).

Stefanidis, Verty, Allen, Owens, Cowley, Oldfield: "The role of thermogenesis in antipsychotic drug-induced weight gain." in: Obesity (Silver Spring, Md.), Vol. 17, Issue 1, pp. 16-24, (2008) (PubMed).

Target Details

Background: Quantitative determination of total protein at 595nm (Bradford method).
Procedure: 2 min.

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. Simple, direct and automation-ready procedures for measuring protein concentration are very desirable. This 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.