Molecular-assisted immunohistochemical optimization

作者：Omar Mohd Feroz Mohd; Huang Ning; Ou Keli; Yu Kun; Putti Thomas C; Jikuya Hiroyuki; Ichikawa Tetsuo; Nishimura Osamu; Tan Patrick; Salto Tellez Manuel^*

来源：Acta Histochemica, 2010, 112(6): 519-528.

DOI：10.1016/j.acthis.2009.05.010

• Immunohistochemistry validation
• Molecular biology
• Molecular diagnostics
• Protein immunolocalization
• Antibody optimization

Immunohistochemistry (IHC) is an essential tool in diagnostic surgical pathology, allowing analysis of protein subcellular localization The use of IHC by different laboratories has lead to inconsistencies in published literature for several antibodies, due to either interpretative (inter-observer venation) or technical reasons These disparities have major implications in both clinical and research settings In this study, we report our experience conducting an IHC optimization of antibodies against five proteins previously identified by proteomic analysis to be breast cancer biomarkers, namely 6PGL (PGLS), CAZ2 (CAPZA2), PA2G4 (EBP1) PSD2 and TKT Large variations in the immunolocalizations and intensities were observed when manipulating the antigen retrieval method and primary antibody incubation concentration However, the use of an independent molecular analysis method provided a clear indication in choosing the appropriate biologically and functionally relevant "staining pattern" Without this latter step, each of these contradictory results would have been a priori "technically acceptable" and would have led to different biological and functional interpretations of these proteins and potentially different applications in a routine pathology setting Thus, we conclude that full validation of immu

• 出版日期2010