Method for detecting activity of lectin of beans
A technology for lectins and beans, which is applied in the field of measuring the activity of bean lectins, can solve the problems of large error in results, difficult operation, high subjectivity, etc., and achieves high stability and accuracy, simple operation method, and high measurement results. accurate effect
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[0038] Example 1
[0039] Determination of Mung Bean Lectin Activity
[0040] (1) Preparation of mung bean lectin solution
[0041] Take 25g of mung bean, crush it and pass it through a 40-mesh sieve, soak in 100ml PBS buffer solution (0.05M, pH7.0, containing 0.15M NaCl), stir at 0℃ for 6h, then centrifuge at 4℃, 16000×g for 15min, Collect the supernatant, soak the resulting precipitate in PBS buffer solution (0.05M, pH7.0, containing 0.15M NaCl), and stir at 0°C for 3 hours, and then centrifuge at 4°C and 16000×g for 15 minutes to collect The two supernatants were combined to obtain mung bean agglutinin solution.
[0042] (2) Preparation of standard red blood cell solution
[0043]Take fresh rabbit blood added with 10% sodium citrate, centrifuge at 1277×g for 15 min, then suspend the collected red blood cell pellet in normal saline, and centrifuge again, repeat this 5 times until the supernatant is no obvious red; Add potassium phosphate saline buffer (pH 8.5, 5mM, 25ml per ml o...
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[0052] Example 2
[0053] Comparison test of different lectin determination methods
[0054] Various measurement methods are as follows:
[0055] (1) Spectrophotometric method: the preparation of mung bean lectin solution and the preparation of standard red blood cell suspension are the same as those of the enzyme labeling method (see Example 1). The mung bean agglutinin solution prepared in Example 1 was serially two-fold diluted and then the following operations were carried out in sequence: 1ml each of the above solution was pipetted into 15 test tubes of 0.5×8cm, and then 1ml of the standard red blood cell solution prepared in Example 1 was added. Mix each test tube with a vortex mixer, then put it into the 0.5cm colorimetric cell with a light path and let it stand for 90-120 minutes, then carefully (minimize shaking) into the spectrophotometer to measure A 620nm . Then calculate according to the following formula
[0056] log x = log...
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