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Biuret Protein Colorimetric Assay Kit– MSE Supplies LLC

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Biuret Protein Colorimetric Assay Kit

SKU: E-BC-K165-S-500

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Biuret Protein Colorimetric Assay Kit

SKU # E-BC-K165-S
Detection Instrument Spectrophotometer (540 nm)
Detection method Colorimetric method


Product Details

Properties

Synonyms TP
Sample Type Serum, plasma, tissue
Sensitivity 0.373 g/L
Detection Range 0.373-80 g/L
Detection Method Colorimetric method
Assay type Quantitative
Assay time 50 min
Precision Average inter-assay CV: 2.600% | Average intra-assay CV: 1.200%
Other instruments required Micropipettor, Incubator
Other reagents required Normal saline (0.9% NaCl), PBS (0.01 M, pH 7.4)
Storage Reagent 3: -20℃, others: 2-8℃
Valid period 12 months


Dilution of Sample

It is recommended to take 2~3 samples with expected large difference to do pre-experiment before formal experiment and dilute the sample according to the result of the pre-experiment and the detection range (0.373-80 g/L).

The recommended dilution factor for different samples is as follows (for reference only):

Sample type Dilution factor
Porcine serum 2-4
Human serum 2-4
5% Mouse liver tissue homogenization 1
10% Epipremnum aureum tissue homogenization 1
5% Mouse heart tissue homogenization 1

 

Note: The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4).

 

Detection Principle

Any compound that contains two -CONH2 in the molecule can react with alkaline copper solution to form a purple complex, which is known as the biuret reaction. Many peptide bonds (-CONH-) in protein molecules can perform this reaction, and the color degree of all kinds of proteins are essentially the same.

 

Kit Components & Storage

Item Component Specification Storage
Reagent 1 Copper Reagent Powder × 1 vial 2-8°C, 12 months
Reagent 2 Alkali Powder × 1 vial 2-8°C, 12 months, shading light
Reagent 3 50 g/L Protein Standard 1.6 mL ×1 vial -20°C, 12 months

 

Note: The reagents must be stored strictly according to the preservation conditions in the above table. The reagents in different kits cannot be mixed with each other.

 

Technical Data:

Parameter:

Intra-assay Precision

Three human serum samples were assayed in replicates of 20 to determine precision within an assay. (CV = Coefficient of Variation)

Parameters Sample 1 Sample 2 Sample 3
Mean (g/L) 2.50 38.50 95.00
%CV 4.2 3.4 4.4


Inter-assay Precision

Three human serum samples were assayed 20 times in duplicate by three operators to determine precision between assays.

Parameters Sample 1 Sample 2 Sample 3
Mean (g/L) 2.50 38.50 95.00
%CV 6.2 7.5 5.8

 

Recovery

Take three samples of high concentration, middle concentration and low concentration to test the samples of each concentration for 6 times parallelly to get the average recovery rate of 98%.

Standard 1 Standard 2 Standard 3
Expected Conc. (g/L) 15 56 72
Observed Conc. (g/L) 15.2 52.6 71.3
Recovery rate (%) 101 94 99

 

Sensitivity

The analytical sensitivity of the assay is 0.58 g/L. This was determined by adding two standard deviations to the mean O.D. obtained when the zero standard was assayed 20 times, and calculating the corresponding concentration.