Kit for detecting calprotectin in human feces and preparation method thereof
A technology of calprotectin and kits, applied in the biological field, can solve problems such as cumbersome operation, susceptibility to environmental interference, and long extraction time by shaking
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Embodiment 1
[0047] The preparation of embodiment 1 kit of the present invention
[0048] Use thermo company EZ- Sμlfo-NHS-LC-Biotin Biotinylation Kit Bovine serum albumin (BSA) is biotinylated, purified by dialysis to obtain the Bi-BSA conjugate and temporarily stored for future use.
[0049] 1. Preparation of the sample pad: soak the sample pad in 0.25-0.5M Tris-HCl buffer solution containing 5-10% trehalose for 5-10 minutes, dry at 37 degrees for 2 hours and store in a sealed manner.
[0050] 2. Preparation of the reaction membrane: Coat the biotin-bovine serum albumin conjugate on the nitrocellulose membrane as the T line, and coat the diluted goat anti-mouse IgG antibody on the nitrocellulose membrane as the C line , dried at 37°C for 16-22h, sealed and stored for later use.
[0051] 3. Preparation of Conjugate Pad 1
[0052] 3.1 Preparation of anti-calprotectin antibody biotinylated conjugates
[0053] use thermo company Sμlfo-NHS-LC-Biotin Biotinylation Kit biotinylated anoth...
Embodiment 2
[0071] Embodiment 2 Preparation of kit of the present invention
[0072] In the preparation of the detection test strip in the kit, the preparation of the binding pad 1 in step 3, the sealing agent used is 2wt% gelatin (w / w), 2wt% NH 2 - 0.5M MES buffer solution of PEG and 0.5wt% trehalose, other steps are the same as in Example 1.
Embodiment 3
[0073] Embodiment 3 Preparation of kit of the present invention
[0074] The formula of feces extraction buffer in this kit is: 4.0M urea, 0.025M CaCl 2 , 0.25M Tris-HCl buffer solution with a pH of 8.0 of 0.5wt% SDS, 10mM EDTA, 0.5M citric acid, 0.05wt% sodium azide and 5wt% BSA, and other steps were the same as in Example 1.
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