Tyrosine phosphorylation profiling via in situ proximity ligation assay.

ABSTRACT:

Background

Tyrosine phosphorylation (pTyr) is an important cancer relevant posttranslational modification since it regulates protein activity and cellular localization. By controlling cell growth and differentiation it plays an important role in tumor development. This paper describes a novel approach for detection and visualization of a panel of pTyr proteins in tumors using in situ proximity ligation assay.

Methods

K562 leukemia cells were treated with tyrosine kinase and/or phosphatase inhibitors to induce differences in pTyr levels and mimic cells with different malignant properties. Cells were then probed with one antibody against the pTyr modification and another probe against the detected protein, resulting in a detectable fluorescent signal once the probes were in pr

SUBMITTER: Elfineh L 

PROVIDER: S-EPMC4072613 | biostudies-literature | 2014 Jun

REPOSITORIES: biostudies-literature