Quantitative determination kit of chemiluminescence immunoassay for Vitamin B12 (VB 12) and preparation method thereof
A chemiluminescence immunoassay and quantitative detection technology, applied in the field of immunoassay medicine, can solve the problems of high cost, narrow detection range, low sensitivity, etc., and achieve the effects of low cost, simple operation and high sensitivity
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Embodiment 1
[0037] Example 1: Preparation of VB12 Chemiluminescent Immunoquantitative Detection Kit
[0038] VB12 chemiluminescent immunoquantitative detection kit, characterized in that the kit includes: VB12 antibody-coated plate, VB12 antigen-enzyme conjugate, VB12 calibrator, VB12 quality control product, 20-fold concentrated lotion, luminescent liquid A liquid and B liquid, sample processing liquid A liquid and B liquid.
[0039] Prepare the VB12 chemiluminescent immunoquantitative detection kit by the following method:
[0040] 1) Preparation of VB12 antibody-coated plates
[0041] Dilute the VB12 antibody with 0.05mol / L pH9.6 carbonate buffer to 1~10ug / mL, add it to a white microwell plate, and coat it at 37°C for 2 hours; discard the liquid in the well, and use pH7.4PBS buffer Wash the plate with liquid, then add 0.5% BSA in phosphate buffer to seal the microwell plate, and seal at 37°C for 2 hours; discard the liquid in the well, spin dry and dry at 37°C for 4 hours; put it int...
Embodiment 2
[0066] Example 2: Preparation of VB12 Chemiluminescent Immunoquantitative Detection Kit
[0067] The kit of the present invention was prepared in the same manner as in Example 1, except that the solid phase carrier was a 48-well white microwell plate.
Embodiment 3
[0068] Embodiment 3: the using method of kit of the present invention
[0069] 1) Equilibrate the test kit at room temperature (18-25°C) for 30 minutes.
[0070] 2) Prepare lotion: Dilute the concentrated lotion with distilled water at 1:20 (1mL lotion plus 19mL distilled water). If the concentrated lotion has crystals, place the concentrated lotion at room temperature or 37°C until the crystals dissolve before diluting.
[0071] 3) Preparation of luminous liquid: 5 minutes before use, take an appropriate amount of luminous liquid A and B and mix them in equal volumes.
[0072] 4) Preparation of sample treatment solution: Prepare sample treatment solution according to the number of test wells. Each well needs 500 μL of sample treatment solution A and 10 μL of sample treatment solution B. The preparation can be increased in an appropriate amount, and it is generally advisable to prepare 10% more. Take an appropriate container to mix the sample treatment solutions A and B. T...
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