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Quantitative protein microarrays for time‐resolved measurements of protein phosphorylation
Author(s) -
Korf Ulrike,
Derdak Sophia,
Tresch Achim,
Henjes Frauke,
Schumacher Sabrina,
Schmidt Christian,
Hahn Bettina,
Lehmann Wolf D.,
Poustka Annemarie,
Beissbarth Tim,
Klingmüller Ursula
Publication year - 2008
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200800112
Subject(s) - proteomics , phosphorylation , dna microarray , protein array analysis , computational biology , protein phosphorylation , protein microarray , quantitative proteomics , chemistry , biology , biochemistry , gene expression , protein kinase a , gene
Abstract The quantitative analysis of signaling networks requires highly sensitive methods for the time‐resolved determination of protein phosphorylation. For this reason, we developed a quantitative protein microarray that monitors the activation of multiple signaling pathways in parallel, and at high temporal resolution. A label‐free sandwich approach was combined with near infrared detection, thus permitting the accurate quantification of low‐level phosphoproteins in limited biological samples corresponding to less than 50 000 cells, and with a very low standard deviation of approximately 5%. The identification of suitable antibody pairs was facilitated by determining their accuracy and dynamic range using our customized software package Quantpro . Thus, we are providing an important tool to generate quantitative data for systems biology approaches, and to drive innovative diagnostic applications.