Kit for detecting concentration of stimulating thyrotrophin receptor antibody
A thyroid-stimulating hormone and detection kit technology is applied in the field of detection kits for stimulating thyroid-stimulating hormone receptor antibody concentration, which can solve the problems of large error in experimental results, long detection time, complicated operation and the like, and achieves a wide detection range, High sensitivity and improved detection efficiency
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preparation example Construction
[0034] In the embodiment of the present invention, the preparation method of the kit of the present invention comprises:
[0035] The first step, the preparation of magnetic particle suspension coated with stimulating thyroid-stimulating hormone receptor antibody
[0036] Take carboxyl magnetic particles according to the amount used, and activate them under acidic conditions with EDC and NHS. After the activation is completed, apply a magnetic field and let it stand to separate the magnetic particles from the liquid. Discard the supernatant and buffer with 0.01M PBS with pH 7.6. Wash away the excess activator with liquid; add an appropriate amount of stimulating thyroid-stimulating hormone receptor antibody, and shake the reaction under weak alkaline conditions; after the reaction, add a magnetic field and let it stand to separate the magnetic particles from the liquid. 0.01M PBS buffer solution of 1% bovine serum albumin for blocking, after the blocking, add blocking solution...
Embodiment 1
[0050] Embodiment 1 reagent preparation
[0051] The reagent preparation method of this embodiment comprises the following steps:
[0052] 1. Preparation of magnetic particle suspension coated with stimulating thyroid-stimulating hormone receptor antibody:
[0053] Take an appropriate amount of carboxyl magnetic particles according to the use, and activate them under acidic conditions (pH4.5-pH5.5) with excess EDC and NHS, and the activation buffer is 0.05M-0.1M MES (2-(N-? pheno)ethanesulfonic acid) buffer solution, the activation time is 30min; after the activation is completed, apply a magnetic field, and let it stand for 5min to separate the magnetic particles from the liquid; discard the supernatant, and wash twice with 0.01M PBS buffer solution of pH 7.6 , to wash away the excess activator; add an appropriate amount of stimulating thyroid stimulating hormone receptor antibody, so that the concentration is 22μg / mg magnetic particles, in 0.05M-0.1M MES buffer (pH4.5-pH5.5...
Embodiment 2
[0060] Embodiment 2 detection method
[0061] 1. Take out a certain amount of reaction containers (reaction wells) and number them, and add 50 μL of clinical serum samples or calibrators to each reaction well according to the experimental requirements;
[0062] 2. Shake the magnetic particle suspension, add 20 μL to each well;
[0063] 3. Add 50 μL of enzyme markers to each well;
[0064] 4. Mix the solution in the reaction well evenly and incubate at 37°C for 30 minutes;
[0065] 5. Using magnetic separation and washing equipment, wash the magnetic particles in the reaction well with washing solution for 5 times;
[0066] 6. Fully shake the washed reaction well to disperse the magnetic particles;
[0067] 7. Add 50 μL each of Luminescent Substrate A and Luminescent Substrate B to each well, shake and mix well, and keep away from light as a reaction for 5 minutes;
[0068] 8. Use a chemiluminescence detection instrument to detect the luminous intensity;
[0069] 9. Adopt ...
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