Enzyme linked immunosorbent assay kit for detecting progesterone and detection method thereof
An enzyme-linked immunosorbent reagent and detection method technology, which is applied in the field of enzyme-linked immunosorbent immunoassay kits for detecting progesterone in chicken tissue, can solve the problem of no progesterone in chicken, and achieve the effect of simple operation and sensitive detection
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[0027] Preparation of progesterone protein conjugates:
[0028] The carboxylated progesterone hapten derivative was obtained by the succinic anhydride method, and then 0.05 mmol was mixed with the carrier protein BSA at a binding ratio of 10:1 in 0.05 mol / L carbonate buffer solution (CBS) at pH 9.6, and then Add 0.15 mmol carbodiimide, stir and react at room temperature for 24 h, and finally dialyze in 0.2 mol / L PBS buffer solution with pH 7.6 for two days to remove unreacted hapten, and put the obtained protein conjugate solution at -20 Store at ℃ for later use.
[0029] Preparation of progesterone antibody:
[0030] Select healthy adult purebred BALA / C mice, take 50 μg of immune antigen prepared by coupling with protein and mix with the same amount of complete Freund's adjuvant, and use intraperitoneal injection for initial immunization, and then use the same dose of immune antigen plus the same amount every 3 weeks Incomplete Freund's adjuvant was used for the second and ...
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