Detection kit for interleukin-18 (IL-18)
A detection kit and interleukin technology, which can be used in measurement devices, instruments, scientific instruments, etc., can solve the problem of low detection sensitivity of interleukin-18 detection
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Embodiment 1
[0035] This embodiment provides a method for preparing an enzyme conjugate, including the following steps:
[0036] 1. Weigh an appropriate amount of Traut's reagent and use antibody activation buffer (100 mM triethanolamine buffer, pH 8.5) to make a 1.376 mg / mL solution.
[0037] 2. Weigh an appropriate amount of Sulfo-SMCC reagent, and use dimethylformamide DMF to prepare a solution with a concentration of 17.5 mg / mL.
[0038] 3. Store the IL-18 labeled antibody in the antibody activation buffer (100mM triethanolamine buffer, pH8.5) at a concentration of 2mg / mL; add the Traut's solution prepared in step 1 to this solution, and IL-18 labeled The molar ratio of antibody to Traut's reagent is 1:15, mix immediately, and let stand at 25°C for 15 minutes.
[0039] 4. Add the Sulfo-SMCC solution prepared in step 2 to the alkaline phosphatase solution (AP solution, the concentration is 20mg / ml), the molar ratio of alkaline phosphatase to Sulfo-SMCC is 1:10, mix immediately, and statically a...
Embodiment 2
[0050] This embodiment provides a method for preparing an enzyme conjugate, including the following steps:
[0051] 1. Weigh an appropriate amount of Traut's reagent and use antibody activation buffer (100 mM triethanolamine buffer, pH 8.5) to make a 1.376 mg / mL solution.
[0052] 2. Weigh an appropriate amount of Sulfo-SMCC reagent, and use dimethylformamide DMF to prepare a solution with a concentration of 17.5 mg / mL.
[0053] 3. Store the IL-18 labeled antibody in antibody activation buffer (100mM triethanolamine buffer, pH 8.5) at a concentration of 5mg / mL; add Traut's solution prepared in step 1 to this solution, and IL-18 labeled The molar ratio of antibody to Traut's reagent is 1:30, mix immediately, and let stand at 25°C for 15 minutes.
[0054] 4. Add the Sulfo-SMCC solution prepared in step 2 to the alkaline phosphatase solution (AP solution, the concentration is 20mg / ml), the molar ratio of alkaline phosphatase to Sulfo-SMCC is 1:15, mix immediately, and statically at 25°C....
Embodiment 3
[0065] This embodiment provides a method for coating magnetic beads, including the following steps:
[0066] 1. Store the IL-18 coated antibody in 100mM MES buffer pH5.0 at a concentration of 1mg / mL;
[0067] 2. Take a carboxyl magnetic bead solution of 100 times the weight of the antibody, the particle size of the carboxyl magnetic bead is 3μm, magnetically separate and discard the supernatant;
[0068] 3. Washing: Re-dissolve the magnetic beads obtained in the above steps with 100mM MES buffer pH5.0, and magnetically separate and discard the supernatant.
[0069] 4. Repeat step 3 once
[0070] 5. Add an appropriate amount of 100mM MES buffer to dissolve the magnetic beads obtained in step 4.
[0071] 6. Weigh an appropriate amount of NHS reagent and dissolve it into a solution with a concentration of 10 mg / mL in 100 mM MES buffer pH 5.0;
[0072] 7. Weigh an appropriate amount of EDC reagent and dissolve it into a solution with a concentration of 10 mg / mL in 100 mM MES buffer pH 5.0;
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