Free triiodothyronine measurement kit and fabrication method thereof
A technology of triiodothyronine and protosine, applied in the field of in vitro testing, can solve the problems of drift of working curve over time, unfavorable automatic quantitative analysis, nonlinear downward shift of low-end slope, etc. Fast, well-performing results
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[0058] The preparation method of solid-phase reagent R1 is:
[0059] A. Activation of paramagnetic microspheres: take paramagnetic microspheres, wash with pH6.0~8.0, 10mM phosphate buffer, resuspend in the above buffer; use concentration of 0.5%~5% EDC solution to activate, in Under the condition of 30-40°C, shake and mix for 30-40 minutes to obtain paramagnetic microsphere resuspension;
[0060] B. Antigen coating: Add the T3 antigen analogue to the resuspension of paramagnetic microspheres, the feeding ratio is 0.2%-2%, and react at 25-37°C for 2-3 hours; then at 25-37°C Blocked for 20-40 minutes under certain conditions; wash the coated paramagnetic microspheres with pH 6.0-8.0, 10mM phosphate buffer, resuspend in the above buffer, and obtain T3 with a concentration of 0.05-0.5mg / ml Suspensions of analogue-coated paramagnetic microspheres;
[0061] The preparation method of liquid phase reagent R2 is:
[0062] Take the T3 antibody, dilute it with pH 9.5, 0.5M / L phosphate...
Embodiment 1
[0070] The preparation of embodiment 1 free triiodothyronine assay kit
[0071] Preparation of solid phase reagent R1:
[0072] A. Activation of paramagnetic microspheres: take the paramagnetic microspheres with -COOH groups on the surface, wash them twice with pH 6.0, 10mM phosphate buffer, remove the supernatant after magnetic separation; add the EDC prepared by the above phosphate buffer (concentration: 0.5%) solution, vortex and mix, and activate at 30°C for 40 minutes; remove the supernatant by magnetic separation, and then add the above-mentioned phosphate buffer solution to wash for 3 times to obtain a paramagnetic microsphere resuspension;
[0073] B. Antigen coating: Add the T3 antigen analogue to the resuspension of paramagnetic microspheres, the feeding ratio is 0.2%, react at 25°C for 3 hours, remove the supernatant by magnetic separation; then add a blocking agent such as containing TRIS buffer solution of protein or amino acid (concentration: 10%), block at 25°C...
Embodiment 2
[0076] The preparation of embodiment 2 free triiodothyronine determination kit
[0077] Preparation of solid phase reagent R1:
[0078] A. Activation of paramagnetic microspheres: take the paramagnetic microspheres with -COOH groups on the surface, wash them twice with pH 8.0, 10mM phosphate buffer, remove the supernatant after magnetic separation; add the EDC prepared by the above phosphate buffer (Concentration: 5%) solution, vortex and mix, activate at 40°C for 30 minutes; remove the supernatant by magnetic separation, then add the above-mentioned phosphate buffer to wash twice, and obtain a paramagnetic microsphere resuspension;
[0079] B. Antigen coating: Add the T3 antigen analogue to the resuspension of paramagnetic microspheres, the feeding ratio is 2%, react at 37°C for 2 hours, remove the supernatant by magnetic separation; then add a blocking agent such as containing TRIS buffer solution of protein or amino acid (concentration: 10%), block at 37°C for 20 minutes; ...
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