Rapid detection and quantification of specific proteins by immunodepletion and microfluidic separation.

ABSTRACT: Conventional immunoblotting techniques are labor intensive, time consuming and rely on the elution of target protein after depletion. Here we describe a new method for detection and quantification of proteins, independent of washing and elution. In this method, the target protein is first captured by immunodepletion with antibody-coated microbeads. In the second step, both the supernatant after immunodepletion and the untreated protein sample are directly analyzed by microfluidic electrophoresis without further processing. Subsequently, the detection and quantification are performed by comparing the electropherograms of these two samples. This method was tested using an Escherichia coli lysate with a FLAG-tagged protein and anti-FLAG magnetic beads. An incubation of as short as one min was

SUBMITTER: Azadi G 

PROVIDER: S-EPMC3517156 | biostudies-literature | 2012 Aug

REPOSITORIES: biostudies-literature