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Merck
CN

Rapid quantification of prion proteins using resistive pulse sensing.

The Analyst (2020-02-18)
Matthew J Healey, Muttuswamy Sivakumaran, Mark Platt
摘要

Prion diseases are a group of fatal transmissible neurological conditions caused by the change in conformation of intrinsic cellular prion protein (PrPC). We present a rapid assay using aptamers and resistive pulse sensing, RPS, to extract and quantify PrPC from complex sample matrices. We functionalise the surface of superparamagnetic beads, SPBs, with a DNA aptamer. First SPB's termed P-beads, are used to pre-concentrate the analyte from a large sample volume. The PrPC protein is then eluted from the P-beads before aptamer modified sensing beads, S-beads, are added. The velocity of the S-beads through the nanopore reveals the concentration of the PrPC protein. The process is done in under an hour and allows the detection of picomol's of protein.

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Sigma-Aldrich
Phosphate buffered saline, tablet
Sigma-Aldrich
Triton X-100, laboratory grade
Sigma-Aldrich
纤维蛋白原 来源于人类血浆, 50-70% protein (≥80% of protein is clottable)
Sigma-Aldrich
白蛋白 来源于人类血清, lyophilized powder, ≥97% (agarose gel electrophoresis)
Sigma-Aldrich
γ-球蛋白 来源于人类血液, ≥99% (electrophoresis)