Biuret Protein Colorimetric Assay Kit
Catalogue Number: E-BC-K165-S-ELA
| Manufacturer: | Elabscience |
| Shelf Life: | 6 months |
| Type: | Protein Detection |
| Shipping Condition: | Blue Ice |
| Unit(s): | 100 assays, 500 assays |
| Range: | 0.373-80 g/L |
| Sensitivity: | 0.373 g/L |
| Sample type: | Plasma, Serum, Tissue |
| Sample size: |
Description
Description: Application The kit can be used to measure total protein (TP) content in serum (plasma), tissue and cultured cells. 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. Experimental instruments Test tube, Micropipettor, Vortex mixer, 37°Cwater bath, Spectrophotometer (540 nm) Preparation of sample It is recommended to take 2~3 samples which expected large difference to do pre-experiment before formal experiment: 1. Serum: detect directly. 2. 10% tissue homogenate: Accurately weigh the tissue, add 9 times the volume of 1xPBS (0.01 M, pH 7.4) according to the ratio of Weight (g): Volume (mL) =1:9. Take the mechanical homogenization in ice water bath. Centrifuge at 10000 g for 10 min, then take 50 µL of the supernatant for detection. Operation procedures 1. Blank tube: Add 50 µL of PBS into a 5 mL EP tube. Standard tube: Add 50 µL of 50 g/L Protein standard into a 5 mL EP tube. Sample tube: Add 50 µL of Sample into a 5 mL EP tube. 2. Add 2.5 mL of biuret working solution into each tube, mix fully with a vortex mixer. 3. Incubate the tubes at 37°C for 30 min, then cool the tubes with running water. 4. Set the spectrophotometer to zero with double-distilled water and measure the absorbance at 540 nm with 1 cm diameter cuvette. Note: It can be refer to the following operating table Technical parameters 1. The sensitivity of the kit is 0.373 g/L. 2. The intra-assay CV is 1.2 % and the inter-assay CV is 2.6%. 3. The recovery of the kit is 99 %. 4. The detection range of the kit is 0.373-80 g/L. Note 1. This kit is for research use only. 2. Instructions should be followed strictly, changes of operation may result in unreliable results. 3. The validity of the kit is 6 months. 4. Do not use components from different batches of kit. 5. If the protein is higher than 80 g/L, please dilute the sample with 1xPBS (0.01 M,pH 7.4) and re-test. If the protein content is lower than 5 g/L, it is suggested to detect the sample with BCA method (E-BC-K318) or Coomassie brilliant blue method (E-BC-K168).
Additional Text
Reagent3:-20°C;Others:2-8°C
E-BC-K165-S Protocol
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