Choice of Fixative Is Crucial to Successful Immunohistochemical Detection of Phosphoproteins in Paraffin-embedded Tumor Tissues

Protein phosphorylation is frequently used as an indicator of cellular signaling activity. Elevated phosphorylation of tyrosine kinase receptors plays an important role in cancer pathogenesis. However, phosphoproteins are usually poorly preserved in clinical tissue samples that are routinely fixed i...

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Published in:The journal of histochemistry and cytochemistry 2009-03, Vol.57 (3), p.257-264
Main Authors: Burns, Janine A, Li, Yuan, Cheney, Carol A, Ou, Yangsi, Franlin-Pfeifer, Laura L, Kuklin, Nelly, Zhang, Zhi-Qiang
Format: Article
Language:English
Subjects:
Animals
Biomarkers, Tumor - analysis
Breast Neoplasms - chemistry
Colonic Neoplasms - chemistry
Data Interpretation, Statistical
Female
Fixatives
Formaldehyde
Humans
Immunohistochemistry
Lung Neoplasms - chemistry
Mice
Mice, Nude
Neoplasm Transplantation
Neoplasms - chemistry
Phosphoproteins - analysis
Polymers
Transplantation, Heterologous
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Summary:Protein phosphorylation is frequently used as an indicator of cellular signaling activity. Elevated phosphorylation of tyrosine kinase receptors plays an important role in cancer pathogenesis. However, phosphoproteins are usually poorly preserved in clinical tissue samples that are routinely fixed in 10% formalin. Nonetheless, in oncology clinical trials, use of phosphoproteins as biomarkers has been considered to be of great value in evaluating the effectiveness of a given drug candidate. Therefore, it is worthy of investigating whether alternative fixatives would improve the preservation of phosphoproteins in tissue. We compared the IHC staining of a number of phosphoproteins in xenograft and human surgical tumor tissues fixed in three different fixatives: 10% formalin, 4% paraformaldehyde (PFA), and Streck's tissue fixative (STF). We found that STF significantly enhanced the staining intensity of phosphoproteins compared with 10% formalin or 4% PFA. STF fixative also showed superiority of preservation of phosphoproteins in human surgical samples. Our results indicate that the choice of fixative could significantly affect the usability of clinical tissue samples for evaluating phosphoprotein by IHC.
ISSN:0022-1554
1551-5044
DOI:10.1369/jhc.2008.952911