A kind of ad biomarker and detection method thereof
A technology of biomarkers and detection methods, applied in the field of AD biomarkers and their detection, can solve the problems of variation, AD biomarkers limiting detection results, etc.
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[0039] (1) Extraction of protein in saliva
[0040] 1. Sample classification, saliva of 15 cases of AD patients and 15 cases of age-matched normal control group were marked.
[0041] 2. Centrifuge at 12000r at 4°C for 10min, and absorb the supernatant.
[0042] 3. Add 450ul supernatant to the column of 10KD ultrafiltration tube, centrifuge at 11000r for 20min, repeat 3-4 times, pay attention to centrifuge the liquid on the column.
[0043] 4. Take 200ul of 50mM Tris-Hcl ((hydroxymethyl)aminomethane) to wash, centrifuge at 11000r for 20min, repeat twice.
[0044] 5. Add 200ul 50mM Tris-Hcl to dissolve the centrifuged protein.
[0045] 6. BCA quantitative determination of protein concentration.
[0046] 7. Take out 100ug protein into a new EP tube (plastic tube), keep the rest, seal and freeze dry.
[0047] (2) Salivary protease digestion step
[0048] 1. According to the protein quantification results, add 100 μg salivary protein to a 1.5 mL centrifuge tube, use Buffer (b...
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