Kit for evaluating semen quality and use method thereof
A sperm quality and kit technology, applied in the field of medical testing, can solve the problems of high price, complicated technical process, high skill and experience requirements for experimental equipment and operators, and achieve the goal of reducing negative impact, simple operation method, and few types of consumables Effect
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Embodiment 1
[0055] A test kit for sperm quality assessment, the test kit includes cell lysate without enzyme activity inhibitors, detection buffer 1, protease inhibitor cocktail, reagent A solution, reagent B solution, bovine serum albumin, fluorescent Reagent, sialidase standard stock solution and detection buffer 2;
[0056] The detection buffer 1 is a phosphate buffer;
[0057] The reagent A liquid is: 1% BCA disodium salt, 2% anhydrous sodium carbonate, 0.16% sodium tartrate, 0.4% sodium hydroxide, 0.95% sodium bicarbonate, mixed to adjust the pH value to 11.2;
[0058] The reagent B liquid is: 4% copper sulfate;
[0059] The detection buffer 2 is a phosphate buffer containing 0.05 mmol / l sodium acetate and 0.25 μg / μl bovine serum albumin.
[0060] Adopt kit of the present invention to measure the sialidase activity of normal sperm (people) and sterile sperm (people) respectively, the result shows that the sialidase activity of normal sperm is obviously higher than that of sterile s...
Embodiment 2
[0062] A test kit for sperm quality assessment, the test kit includes cell lysate without enzyme activity inhibitors, detection buffer 1, protease inhibitor cocktail, reagent A solution, reagent B solution, bovine serum albumin, fluorescent Reagent, sialidase standard stock solution and detection buffer 2;
[0063] The detection buffer 1 is a phosphate buffer;
[0064] The reagent A liquid is: 1% BCA disodium salt, 2% anhydrous sodium carbonate, 0.16% sodium tartrate, 0.4% sodium hydroxide, 0.95% sodium bicarbonate, mixed to adjust the pH value to 11.3;
[0065] The reagent B liquid is: 4% copper sulfate;
[0066] The detection buffer 2 is a phosphate buffer containing 0.05 mmol / l sodium acetate and 0.25 μg / μl bovine serum albumin.
[0067] The sialidase standard stock solution is Clostridium perfringens sialidase with a concentration of 5 U / ml.
Embodiment 3
[0069] A test kit for sperm quality assessment, the test kit includes cell lysate without enzyme activity inhibitors, detection buffer 1, protease inhibitor cocktail, reagent A solution, reagent B solution, bovine serum albumin, fluorescent Reagent, sialidase standard stock solution and detection buffer 2;
[0070] The detection buffer 1 is BWW culture fluid;
[0071] The reagent A liquid is: 1% BCA disodium salt, 2% anhydrous sodium carbonate, 0.16% sodium tartrate, 0.4% sodium hydroxide, 0.95% sodium bicarbonate, mixed to adjust the pH value to 11.25;
[0072] The reagent B liquid is: 4% copper sulfate;
[0073] The detection buffer 2 is a phosphate buffer containing 0.05 mmol / l sodium acetate and 0.25 μg / μl bovine serum albumin.
[0074] The sialidase standard stock solution is Clostridium perfringens sialidase with a concentration of 5 U / ml.
[0075] The fluorescent reagent is sodium 2'-4-methylumbelliferyl-α-D-N-acetylneuraminic acid.
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