1. Drawing of standard curve
(1) Prepare standard bovine serum albumin solution: accurately weigh 0.025g of crystallized bovine serum albumin on the analytical balance, pour it into a small beaker, add a small amount of distilled water to dissolve it, and then transfer it to 100m 1
In the volumetric flask, the residual liquid in the beaker is washed several times with a small amount of distilled water, and the washing liquid is poured into the volumetric flask together. Finally, the volume is adjusted to the scale with distilled water, and a standard egg autotetrate solution is prepared, in which the concentration of bovine serum albumin is 250 ug/ ml.
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(2) Preparation of serial standard bovine serum albumin solutions: Take 6 test tubes with stoppers, and add bovine serum albumin standard solutions and distilled water according to the following table. Then add 5ml of reagent A to each tube, mix them and leave them at room temperature.
10min, add 0.5ml of reagent B, and immediately mix evenly (this step is faster, otherwise the color development degree will be weakened). After 30min, the protein-free 1 test tube was used as the control, compared with other 5.
The solution in the test tube is colorized by spectrophotometer at the wavelength of 5OOnm in turn. Record the absorbance of the solution in each test tube.
Reagent 1 2 3 4 5 6
250ug/mg bovine serum protein (ml) 00.20.40.60.810.
H 2 O (ml) 1 0.8 0.6 0.4 0.2 0
Protein content (ug) 0 50/kloc-0 00/kloc-0 50 200 250.
(3) Drawing of standard curve: Drawing a standard curve with absorbance as the vertical scale and bovine serum albumin content (ug) as the horizontal scale.
2. Sample determination
(1) Weigh hypocotyl of mung bean sprouts 1g into a mortar, add distilled water 2m 1 and homogenize. Transfer to a centrifuge tube, wash the mortar with 6m 1 distilled water for several times, and merge into the centrifuge tube. Centrifuge at 4000r/min for 20min. Discard the precipitate, transfer the supernatant to a volumetric flask, and fix the volume to 10m 1.
(2) Take 2 test tubes with stoppers, add 1 ml of supernatant to each tube, add 5m 1 of reagent A respectively, mix well, place lomin, then add 0.5 ml of reagent B, mix well quickly, place at room temperature for 30min, compare colors at the wavelength of 500nm, and record the absorbance value.