Phenol reagents determine proteins

related topicsPrinciple:Protein
and alkaline copper solution action (double shrink reaction), the protein in the copper-linked protein contains tyrosine and tryptophan residues, so that phosphate - phosphorus tungsten acid (Fotin) -Ciocalteau phenol
reagement
) reduction is blue, the shade of blue is directly related to the content of aromatic
amino acids
so it can be compared to the color method.Operations1. The drawing of the standard curve:take seven
dring

test tubes
in the number, operate according to the table:mix well, place for 10 minutes, 0.5mL per tube of phenolic reagents, quickly mix well. After 30 minutes, the 6th tube is used as a blank tube, at a wavelength of 650nm color, to read the light density.
1-5 tube protein concentration as the horizontal coordinates, light density as ordinates, to map the standard curve. After drawing, indicate the instrument model, number, wavelength used, measurement method and production date.
according to the light density of the 7th tube, the corresponding concentration is found by the standard curve. The concentration unit of the sample is converted into protein (g)/100mL
serum
.
Note:
1. After treatment with alkaline copper solution, protein color ability increased by 3 to 15 times. Copper treatment has less effect on the hair color ability of tyrosine and tryptophan, and 1/4 of the colors shown in this method are derived from the double-shrink reaction.
2. The content of protein in the sample should be within 300 μg/mL, if the concentration exceeds this range, the reaction is not linear.
3. Hair color reaction in 30 minutes is close to the limit, in half an hour to an hour the color slightly increased, in 1.5 to 6 hours the color is stable, therefore, it is best to color within 1.5 to 6 hours after the color.

1.15×15cm test tube
2.scale
straw
(0.5mL, 1mL, 5mL)
3.721
dlight meter 4.coordinate meter
5.micro-addition Device (500 sil)
reagentsstandard bovine serum protein solution: accurately called 0.4g of bovine serum protein, dissolved in 100mL0.9% NaCl solution, diluted 10 times used.