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SDS-PAGE/Immunoblot Detection of Aβ Multimers in Human Cortical Tissue Homogenates using Antigen-Epitope Retrieval
Published on: April 23, 2010
Quantum dots-based sandwich immunoassay for sensitive detection of Alzheimer's disease-related Aβ1-42
Menghuan Tang1, Jiangli Pi1, Yijuan Long1
1Key Laboratory of Luminescent and Real-Time Analytical Chemistry (Southwest University), Ministry of Education, College of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715, PR China.
Abstract:
Amyloid-beta peptide 1-42 (Aβ1-42) is known as a component of amyloid plaques in association with Alzheimer's disease. Herein, we developed a reliable and remarkably sensitive sandwich immunoassay to detect the Aβ1-42 using quantum dots (QDs) as fluorescent label. In the presence of Aβ1-42, the biotinylated Anti-beta Amyloid 1-16 (N-Ab) recognized the target and formed C-Ab-Aβ1-42-N-Ab sandwich immunocomplexes. Then Streptavidin-QDs conjugated to biotinylated N-Ab and the concentration of Aβ1-42 was determined by detecting the fluorescence intensity in the supernatant. This method is faster and more efficient than the previous approach we reported. It also has reasonable sensitivity and selectivity. Under the optimized conditions, the linear range is 5.0 to 100 pM (0.023-0.45 ng/mL) and the detection limit is 1.7 pM (7.6 pg/ mL). In addition, this method has been successfully applied to detect the Aβ1-42 in human cerebrospinal fluid sample.

