A reverse phase protein array based phospho-antibody characterization approach and its applicability for clinical derived tissue specimens.

ABSTRACT: Systematic quantification of phosphoprotein within cell signaling networks in solid tissues remains challenging and precise quantification in large scale samples has great potential for biomarker identification and validation. We developed a reverse phase protein array (RPPA) based phosphor-antibody characterization approach by taking advantage of the lysis buffer compatible with alkaline phosphatase (AP) treatment that differs from the conventional RPPA antibody validation procedure and applied it onto fresh frozen (FF) and formalin-fixed and paraffin-embedded tissue (FFPE) to test its applicability. By screening 106 phospho-antibodies using RPPA, we demonstrated that AP treatment could serve as an independent factor to be adopted for rapid phospho-antibody selection. We also showed desir

SUBMITTER: Wang N 

PROVIDER: S-EPMC9792559 | biostudies-literature | 2022 Dec

REPOSITORIES: biostudies-literature