Jump to main content
Jump to site search
Access to RSC content Close the message box

Continue to access RSC content when you are not at your institution. Follow our step-by-step guide.


Issue 13, 2014
Previous Article Next Article

Absolute quantification of protein copy number using a single-molecule-sensitive microarray

Author affiliations

Abstract

We report the use of a microfluidic microarray incorporating single molecule detection for the absolute quantification of protein copy number in solution. In this paper we demonstrate protocols which enable calibration free detection for two protein detection assays. An EGFP protein assay has a limit of detection of <30 EGFP proteins in a microfluidic analysis chamber (limited by non-specific background binding), with a measured limit of linearity of approximately 6 × 106 molecules of analyte in the analysis chamber and a dynamic range of >5 orders of magnitude in protein concentration. An antibody sandwich assay was used to detect unlabelled human tumour suppressor protein p53 with a limit of detection of approximately 21 p53 proteins and a dynamic range of >3 orders of magnitude. We show that these protocols can be used to calibrate data retrospectively to determine the absolute protein copy number at the single cell level in two human cancer cell lines.

Graphical abstract: Absolute quantification of protein copy number using a single-molecule-sensitive microarray

Back to tab navigation

Article information


Submitted
13 Jan 2014
Accepted
11 Mar 2014
First published
11 Mar 2014

Analyst, 2014,139, 3235-3244
Article type
Paper

Absolute quantification of protein copy number using a single-molecule-sensitive microarray

E. Burgin, A. Salehi-Reyhani, M. Barclay, A. Brown, J. Kaplinsky, M. Novakova, M. A. A. Neil, O. Ces, K. R. Willison and D. R. Klug, Analyst, 2014, 139, 3235
DOI: 10.1039/C4AN00091A

Social activity

Search articles by author

Spotlight

Advertisements