Kit for enzyme-linked immunosorbent assay of cordyceps sinensis and preparation method of kit
An enzyme-linked immunoassay and cordyceps sinensis technology, which is applied in the direction of biological testing, measuring devices, material inspection products, etc., can solve the problems of inability to identify, weight gain, etc., and achieve the effects of wide application range, good accuracy and high sensitivity
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Embodiment 1
[0021] Example 1 Preparation of target antigen.
[0022] After grinding the body part of Cordyceps sinensis under liquid nitrogen conditions, 10 g of the sample was taken, and 100 ml of 0.1M PBS buffer (pH7.4, containing 0.5% mercaptoethanol, 0.1% Tween80) was ultrasonically extracted for 12 hours in an ice bath. The supernatant was collected by centrifugation and precipitated with 60% saturation ammonium sulfate. 8M urea was used to dissolve the precipitate, centrifuged, and the supernatant was dialyzed overnight to obtain a crudely separated sample.
[0023] The primary separated sample was repurified using a vertical electrophoresis apparatus: the primary separated sample was subjected to SDS-PAGE electrophoresis using a 4% stacking gel and a 12% separating gel, and a prestained protein standard was used as a molecular weight reference. After electrophoresis at 80v for 2 hours, a protein gel band with a molecular weight of 70kd was cut out with reference to the prestained ...
Embodiment 2
[0024] Example 2 Preparation and purification of anti-Cordyceps sinensis antibody.
[0025] The immunogen prepared in Example 1 was used to immunize female balb / c mice for 6 weeks respectively, 3 mice in each group. For the first immunization injection, 100 μL of 100 μg / mL immune antigen was fully emulsified with the same amount of complete Freund’s adjuvant, and injected directly into the intraperitoneal cavity. After an interval of two weeks, take the sampled antigen, emulsify it with 100 μL of incomplete adjuvant, and inject it in the same way.
[0026] One day before cell fusion or on the same day, the Kunming rats were killed by pulling the neck, soaked in 70% alcohol, and disinfected the body surface; the Kunming rats were fixed on the wax board with pins, the abdomen was cut open on the ultra-clean workbench, and the peritoneum was picked up with small tweezers. Inject 5 mL of RPMI-1640 complete culture solution (obtained by adding 15% fetal calf serum to GIBICO RPMI-1...
Embodiment 3
[0033] Example 3 Preparation of standard substance of Cordyceps sinensis.
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