Immuno-digital invasive cleavage assay for analyzing Alzheimer’s amyloid ß-bound extracellular vesicles - Alzheimer's Research & Therapy

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Immuno-digital invasive cleavage assay for analyzing Alzheimer’s amyloid ß-bound extracellular vesicles - Alzheimer's Research & Therapy
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An article published in Alzheimer's Research & Therapy presents a method for the quantitative detection of extracellular vesicles as a blood biomarker for preclinical Alzheimer’s disease diagnosis.

for 1 h to pellet the EVs. EV protein content was measured with the BCA assay . Sizes and densities of the EVs suspended in PBS were analyzed with a qNano system using NP200 nanopores and qNano Izon analysis software. CPC100 was used as the calibration sample in this study. Microphotographs of the EVs were obtained with an HD-2000 scanning transmission electron microscope .

5 μg CTB at room temperature for 30 min with rotation. Then, 10 μL of 10 mg/mL 1-ethyl-3-carbodiimide hydrochloride in 0.1 M MES buffer was added into the mixture and rotated at room temperature for 3 h. CTB-coupled beads were then blocked with 0.5% bovine serum albumin , washed twice with PBST , and resuspended in 100 μL of 0.1% Tween-20/PBS until use.

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