Calprotectin chemiluminescence immunoassay kit and preparation method thereof
A technology of chemiluminescent immunity and detection kits, which is applied in biological testing, measuring devices, scientific instruments, etc., and can solve the problems of long operation time, only qualitative, low degree of automation, etc.
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[0045] The preparation method of the calprotectin chemiluminescent immunoassay kit of the present invention mainly comprises the following steps:
[0046] Step 1. Preparation of streptavidin magnetic particles
[0047] Take streptavidin magnetic particle solution, add TBST solution, mix thoroughly, place on a magnetic separator until the supernatant is free of turbidity, discard the supernatant, and keep the magnetic particles; repeat washing 3 times, wash with 50-100mM MES , 0.05-0.15% Tween-20, 0.05-0.1% Proclin300, resuspended in buffer solution with pH 6.5-7.5;
[0048] Step 2. Preparation of calprotectin antibody labeled with chemiluminescence marker
[0049] Replace the calprotectin antibody buffer with PBS, add a chemiluminescent marker, mix well, and purify the labeled calprotectin antibody after standing at room temperature for 2 to 4 hours; place the collected chemiluminescent marker-labeled calprotectin antibody in Store at 2-8°C; use 50-100mM MES or PBS, 0.05% Twee...
Embodiment 1
[0056] Preparation of embodiment 1 calprotectin chemiluminescent immunoassay kit
[0057] (1) Preparation process of streptavidin magnetic particles
[0058] Take 1 mL of streptavidin magnetic particle solution with Tosyl functional group at a concentration of 100 mg / mL (streptavidin magnetic beads were purchased from Agilent Technologies Co., Ltd.), add 5 mL of TBST solution and mix well for 10 min, then place in On a magnetic separator, until the supernatant is free of turbidity, discard the supernatant, and keep the magnetic particles; after repeated washing 3 times, resuspend with 50mM MES, 0.15% Tween-20, 0.10% Proclin300, pH6.5 buffer to prepare The solid-phase reagent with a final concentration of 0.1% into magnetic beads is the R1 reagent.
[0059] (2) Preparation process of calprotectin antibody labeled with chemiluminescence marker
[0060] Use an ultrafiltration centrifuge tube to replace the calprotectin antibody buffer with PBS (the mass of calprotectin antibody...
Embodiment 2
[0063] Example 2 Preparation of Calprotectin Chemiluminescent Immunoassay Kit
[0064] (1) Preparation process of streptavidin magnetic particles
[0065] Take 1 mL of streptavidin magnetic particle solution with Tosyl functional group at a concentration of 100 mg / mL (streptavidin magnetic beads were purchased from Agilent Technologies Co., Ltd.), add 8 mL of TBST solution and mix well for 15 min, then place in On a magnetic separator, until the supernatant is free of turbidity, discard the supernatant, and keep the magnetic particles; after repeated washing 3 times, resuspend with 75mM MES, 0.10% Tween-20, 0.05% Proclin300, pH7.0 buffer to prepare The solid-phase reagent with a final concentration of 0.05% into magnetic beads is the R1 reagent.
[0066] (2) Preparation process of calprotectin antibody labeled with chemiluminescence marker
[0067] Use an ultrafiltration centrifuge tube to replace the calprotectin antibody buffer with PBS (the mass of calprotectin antibody i...
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