Enhancing single-cell western blotting sensitivity using diffusive analyte blotting and antibody conjugate amplification.

ABSTRACT: While there are many techniques to achieve highly sensitive, multiplex detection of RNA and DNA from single cells, detecting protein contents often suffers from low limits of detection and throughput. Miniaturized, high-sensitivity western blots on single cells (scWesterns) are attractive since they do not require advanced instrumentation. By physically separating analytes, scWesterns also uniquely mitigate limitations to target protein multiplexing posed by affinity reagent performance. However, a fundamental limitation of scWesterns is their limited sensitivity for detecting low-abundance proteins, which arises from transport barriers posed by the separation gel against detection species. Here we address sensitivity by decoupling the electrophoretic separation medium from the detection m

SUBMITTER: Long MA 

PROVIDER: S-EPMC10312704 | biostudies-literature | 2023 Jun

REPOSITORIES: biostudies-literature