Kit for quantitative determination of active oxygen materials in sperms as well as application and using method of kit
A detection kit and oxygen quantification technology, applied in the direction of color/spectral characteristic measurement, measuring device, and analysis through chemical reaction of materials, etc. Convenience and other issues, achieve high sensitivity, overcome imprecise detection, and simple operation
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Embodiment 1
[0050]1. Prepare the quantitative detection kit for sperm reactive oxygen species in Example 1, as follows.
[0051] Prepare NBT freeze-dried powder (0.212mmol / L): Accurately weigh 1.2134g of NBT dry powder, make up to 1000mL with purified water, dispense 1mL / bottle, freeze-dry, and add 7mL NBT solution to redissolve when used, that is, the concentration of NBT is 0.212mmol / L.
[0052] Prepare NBT solution (phosphate buffer solution with a concentration of 0.05mol / L): Weigh 2.5857g of potassium dihydrogen phosphate and 29.0093g of disodium hydrogen phosphate dodecahydrate, add an appropriate amount of purified water and stir to dissolve, then dilute the purified water to 1000mL, adjust the phosphate pH to 7.2-7.6.
[0053] Prepare color developer A (0.25mol / L sodium bicarbonate): weigh 21.0025g of sodium bicarbonate, add purified water to the volumetric flask to make up to 1000mL.
[0054] Prepare color developer B (1.62% triethanolamine): take 16.2 mL of triethanolamine, ad...
Embodiment 2
[0084] 1. Preparation of the quantitative detection kit for sperm reactive oxygen species in Example 2, as follows.
[0085] Preparation of freeze-dried powder (0.1mmol / L NBT): Accurately weigh 0.2862g of NBT dry powder, make up to 500mL with purified water, dispense 1mL / bottle, freeze-dry, and add 7mL NBT solution to redissolve before use, that is, the concentration of NBT is 0.1mmol / L.
[0086] Preparation of NBT solution (phosphate buffer solution with a concentration of 0.01mol / L): Weigh 0.25857g of potassium dihydrogen phosphate and 2.90093g of disodium hydrogen phosphate dodecahydrate, add an appropriate amount of purified water and stir to dissolve, then dilute the purified water to 1000mL, use concentrated hydrochloric acid or concentrated sodium hydroxide solution to adjust the pH to 7.2-7.6.
[0087] Prepare color developer A (0.01mol / L sodium bicarbonate): Weigh 0.8401g of sodium bicarbonate, add purified water to the volumetric flask to make up to 1000mL.
[0088...
Embodiment 3
[0118] 1. Preparation of the quantitative detection kit for sperm reactive oxygen species in Example 3, as follows.
[0119] Prepare NBT freeze-dried powder (1mmol / L): Accurately weigh 5.7236g of NBT dry powder, make up to 1000mL with purified water, dispense 1mL / bottle, freeze-dry, and add 7mL NBT solution to redissolve (that is, the concentration of NBT is 1mmol / L.
[0120] Preparation of NBT solution (phosphate buffer solution with a concentration of 0.5mol / L): Weigh 12.9285 g of potassium dihydrogen phosphate and 145.0465 g of disodium hydrogen phosphate dodecahydrate, add an appropriate amount of purified water and stir to dissolve, then dilute the purified water to 1000mL, use concentrated hydrochloric acid or concentrated sodium hydroxide solution to adjust the pH to 7.2-7.6.
[0121] Prepare color developer A (0.3mol / L sodium bicarbonate): weigh 12.6046g sodium bicarbonate, add purified water to the volumetric flask to make up to 500mL.
[0122] Prepare color develop...
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