Myoglobin detection reagent kit and method for applying same
A technology for detecting kits and myoglobin, applied in the field of myoglobin detection kits, can solve the problems of narrow detection range, low specificity of myoglobin detection kits, cumbersome operation, etc., and achieve detection sensitivity and precision Improve the effect of avoiding non-specific binding and simplifying the operation steps
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Embodiment 1
[0055] The preparation of embodiment 1 enzyme conjugate
[0056] 1. Weigh 3mg of Traut's reagent and prepare a solution with a concentration of 1.376mg / mL with 100mmol / L triethanolamine buffer solution (pH=8.5±0.05); L of triethanolamine buffer solution (pH = 8.5 ± 0.05) was prepared into a solution with a concentration of 2 mg / mL, and was added with a concentration of 1.376 mg / mL of Traut's solution, the mixture of myoglobin-labeled antibody and Traut's reagent The molar ratio is 1:15, mix immediately, and react at room temperature for 15 minutes. Add the glycine solution of 1mmol / L, the amount of substance of glycine is 10 of the amount of 4-(N-maleimidomethyl) cyclohexane-1-carboxylic acid sulfosuccinimide ester sodium salt substance times, mix immediately, and react at room temperature for 10 minutes. Desalted and replaced with 100 mmol / L triethanolamine cross-linking buffer (pH=7.3±0.05).
[0057] 2. Weigh 3 mg of 4-(N-maleimidomethyl) cyclohexane-1-carboxylic acid sul...
Embodiment 2
[0061] The preparation of embodiment 2 enzyme conjugates
[0062] 1. Weigh 3mg of Traut's reagent and prepare a solution with a concentration of 1.3mg / mL with 100mmol / L triethanolamine buffer solution (pH=8.5±0.05); L of triethanolamine buffer solution (pH=8.5 ± 0.05) was prepared into a solution with a concentration of 5 mg / mL, and was added with a concentration of 1.3 mg / mL Traut's solution, the mixture of myoglobin-labeled antibody and Traut's reagent The molar ratio is 1:10, mix immediately, and react at room temperature for 12 minutes. Add the glycine solution of 1mmol / L, the amount of substance of glycine is 20% of the amount of 4-(N-maleimidomethyl) cyclohexane-1-carboxylic acid sulfosuccinimide ester sodium salt substance times, mix immediately, and react at room temperature for 12 minutes. Desalted and replaced with 100 mmol / L triethanolamine cross-linking buffer (pH=7.3±0.05).
[0063] 2. Weigh 3 mg of 4-(N-maleimidomethyl) cyclohexane-1-carboxylic acid sulfosucci...
Embodiment 3
[0067] The preparation of embodiment 3 enzyme conjugates
[0068] 1. Weigh 3mg of Traut's reagent and prepare a solution with a concentration of 1.5mg / mL with 100mmol / L triethanolamine buffer solution (pH=8.5±0.05); L of triethanolamine buffer solution (pH = 8.5 ± 0.05) was prepared into a solution with a concentration of 2.5 mg / mL, and to it was added a concentration of 1.5 mg / mL Traut's solution, myoglobin-labeled antibody and Traut's reagent The molar ratio is 1:15, mix immediately, and react at room temperature for 18 minutes. Add the glycine solution of 1mmol / L, the amount of substance of glycine is 15% of the amount of 4-(N-maleimidomethyl) cyclohexane-1-carboxylic acid sulfosuccinimide ester sodium salt substance times, mix immediately, and react at room temperature for 10 minutes. Desalted and replaced with 100 mmol / L triethanolamine cross-linking buffer (pH=7.3±0.05).
[0069] 2. Weigh 3 mg of 4-(N-maleimidomethyl) cyclohexane-1-carboxylic acid sulfosuccinimide sodiu...
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