Elabscience
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1th Edition, revised in MAY, 2015
（FOR RESEARCH USE ONLY. DO NOT USE IT IN CLINICAL DIAGNOSIS !）
Total protein quantitative assay kit (With standard: Biuret method)
Catalog No: BC0165
Specification： 100 tubes /96 samples
Detection Method：Biuret method
This manual must be read attentively and completely be
fore using this product.
May you have any problems, please contact our Technical Service Center for help.
Phone: 86-27-87385095
Email: [email protected]
[email protected]
Website: www.elabscience.com
Please kindly provide us the lot number (on the outside of the box) of the kit for more efficient service.
1th Edition, revised in MAY, 2015
Product Introduction
The determination of protein concentration is an essential technique in all aspects of protein studies and
proteomics. It bases on Lambert-Beer law, namely there is a linear relationship between absorbance
and concentration of the solution within a certain range of concentration and wavelength. The
kit provides microscale and quick method for determining protein concentration, the operation is
simple, time-saving, and can save a large amount of reagent, and a large number of samples can be measured. Any
compound molecule contains two carbamoyl (-CONH2) is able to react with alkaline copper solution, forming a
purple complex, this reaction is referred to biuret reaction. There are a lot of peptide bonds (CONH) in the protein
molecule can occur in this reaction, A variety of protein have almost the same color degree.
Experimental Instrument
test tube, micropipettor, vortex mixer, 37 ℃ water bath, visible spectrophotometer (540nm)
3 Tested Application
Serum, plasma, tissue and other biological samples.
4 Operation Steps
(1) Sample Preparation
①Serum（plasma）: 50μl samples were measured directly.
②Tissue homogenates: Tissue pieces should be weighed , and add appropriate amount of precooling PBS (the
volume depends on the weight of the tissue. 9mL PBS would be appropriate to 1 gram tissue pieces), then
homogenize on ice to be 10% homogenate. The homogenates are centrifugated for 10 minutes at 2500rpm/min.
Take 50μl supernatant to measured.
(2) Operating Table
Blank tube
Distilled water (ml)
Standard tube
0.05
standard solution(ml)
0.05
Test samples(ml)
Biuret reagent (ml)
Sample tube
0.05
2.5
2.5
2.5
Mixing, water bath at 37℃ for 10 minutes, water cooling, 540nm wavelength, 1cm diameter, zero with
distilled water, measuring the OD values of each tube.
5 Computational formula:
Concentration of protein (mgprot/ml)=
OD s a m p le − OD b la n k
OD s ta n d a r d −OD b la n k
× Concentration of standard (mgprot/ml)
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1th Edition, revised in MAY, 2015
6 Product Advantages
①The operation is simple: single-agent operation, it can measure above 50 samples in 20 minutes.
②Good stability: it can be stored for six months at 2 - 8℃, the color is stable for over 1 hour.
③Good reproducibility: Intra-assay CV = 2.3%, Inter-assay CV = 5.34%.
④Recovery test: X = 100%.
⑤Less affected by external factors: less interference factors and good repeatability.
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