Human Legumain chemiluminiscence detection kit and application thereof
A chemiluminescence detection and kit technology, which is applied in the field of immunodiagnosis, can solve the problems that the detection results are easily affected by the detection conditions, the detection of the kit is not sensitive, and the accuracy is not high, so as to improve the specificity and detection speed, improve the sensitivity and Applicability, effect of increasing luminosity
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[0036] A method for preparing a human Legumain chemiluminescence detection kit, the method comprising:
[0037] Step 1: Preparation of streptavidin-based magnetic bead suspension
[0038] 1) After dissolving and mixing ferric chloride and ferrous chloride in water, add ammonia water and react at 50-70°C for 3-5 hours to obtain magnetic microspheres; the molar ratio of ferric chloride and ferrous chloride The ratio is preferably 2:1;
[0039] 2) Wash the magnetic microspheres obtained in step 1) and redisperse them in 50% ethanol, and then add 0.5% PEG4000, 0.1% ammonia and 0.1% ethyl orthosilicate in sequence at 60-65°C and stir for 14h-18h to obtain Modified magnetic microspheres; the volume ratio of PEG4000, ammonia and ethyl orthosilicate is preferably 1:5:1;
[0040] 3) Wash the modified magnetic microspheres obtained in step 2) alternately with ultrapure water and absolute ethanol, then resuspend in 50% ethanol and add 0.05% ammonia and 0.5% tetramethylammonium hydroxide respect...
Embodiment 1
[0050] (1) Preparation of streptavidin-based magnetic bead suspension
[0051] 1. Weigh ferric chloride and ferrous chloride at a molar ratio of 2:1, dissolve and mix them in ultrapure water, add ammonia to a final concentration of 0.5%, stir and react at 60°C for 4 hours to obtain magnetic microspheres;
[0052] 2. Wash the obtained magnetic microspheres with ultrapure water and redisperse them in 50% ethanol, then add 1mL 0.5% PEG4000, 5mL 0.1% ammonia water and 1mL 0.1% ethyl orthosilicate, and stir at 60℃. React for 16 hours to obtain modified magnetic microspheres;
[0053] 3. Wash the modified magnetic microspheres with ultrapure water and absolute ethanol alternately, then resuspend them in 50% ethanol, and then add 5mL 0.05% ammonia, 2mL 0.5% tetramethylammonium hydroxide TMA and 4mL 2.5% 3-aminopropyltrimethoxysilane, stirred and reacted at room temperature for 16h, to obtain magnetic microspheres with surface modified amino groups;
[0054] Take 8mg of the amino magnetic mi...
Embodiment 2
[0063] (1) Preparation of streptavidin-based magnetic bead suspension
[0064] Weigh ferric chloride and ferrous chloride at a molar ratio of 2:1, dissolve and mix them in ultrapure water, add ammonia water to a final concentration of 0.5%, stir and react at 60°C for 3 hours to obtain magnetic microspheres;
[0065] The obtained magnetic microspheres were washed with ultrapure water and re-dispersed in 50% ethanol, and then 1mL 0.5% PEG4000, 5mL 0.1% ammonia water and 1mL 0.1% ethyl orthosilicate were added successively, and the reaction was stirred at 60°C for 14h. Obtain modified magnetic microspheres;
[0066] The modified magnetic microspheres were washed alternately with ultrapure water and absolute ethanol, then resuspended in 50% ethanol, and then added 5mL 0.05% ammonia, 2mL 0.5% tetramethylammonium hydroxide TMA and 4mL 2.5 % 3-aminopropyltrimethoxysilane, stirred and reacted at room temperature for 14 hours, to obtain magnetic microspheres with surface modified amino group...
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