Journal article
Improved immunohistochemical detection of type 1 insulin-Like growth factor receptor in human tumors
- Abstract:
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Background: Insulin-like growth factors (IGFs) are known to play important roles in cancer biology, prompting evaluation of drugs targeting type 1 IGF receptor (IGF-1R). However, there is considerable lack of consensus in immunohistochemical (IHC) studies of IGF-1R in human tumors, confounding attempts to assess the predictive and prognostic significance of IGF-1R expression and subcellular localization. Likely sources of variation include use of different IGF-1R polyclonal antibodies and methods for IHC. Here, we aimed to develop a robust IGF-1R IHC protocol using a monoclonal antibody, suitable for use in formalin-fixed paraffin-embedded (FFPE) tissues.
Methods: Using controls including samples of FFPE tissues and tumor cells of defined IGF-1R expression, we used IHC and western blotting to compare polyclonal antibody #3027 with monoclonals #9750 and #14534 (Cell Signaling Technology).
Results: Compared with #3027, the monoclonals exhibited superior discrimination between IGF-1R-high and IGF-1R-deficient cells in manual IHC, signal generated by #9750 reflecting differences in IGF-1R expression detected by western blotting. In tissues, IGF-1R detected by #14534 was predominantly plasma membrane-associated, while #9750 detected IGF-1R in the plasma membranes, cytoplasm and nucleus of prostate and renal cancers, recapitulating appearances we described using previous lots of #3027, and reflecting subcellular localizations reported using other techniques. Use of #9750 and #14534 in an autostainer showed adequate differentiation of high vs low IGF-1R cells, but did not recapitulate appearances of manually-stained tissues. We provide a detailed protocol for the preferred manual method using #9750.
Conclusion: Standardization of IGF-1R IHC will promote understanding of the role of IGF-1R in tumor biology, and its potential as a candidate prognostic and predictive biomarker.
- Publication status:
- Published
- Peer review status:
- Peer reviewed
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(Preview, Version of record, pdf, 786.0KB, Terms of use)
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- Publisher copy:
- 10.4172/2469-9756.1000114
Authors
- Publisher:
- OMICS International Journals
- Journal:
- Immunochemistry and Immunopathology More from this journal
- Volume:
- 2
- Issue:
- 1
- Article number:
- 1000114
- Publication date:
- 2016-01-25
- Acceptance date:
- 2016-01-21
- DOI:
- ISSN:
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2469-9756
- Keywords:
- Pubs id:
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pubs:599052
- UUID:
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uuid:92b7d9e8-c87e-4834-be0d-e9b63bbb0b4c
- Local pid:
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pubs:599052
- Source identifiers:
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599052
- Deposit date:
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2016-02-18
Terms of use
- Copyright holder:
- Aleksic et al
- Copyright date:
- 2016
- Notes:
- © 2016 Aleksic T, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
- Licence:
- CC Attribution (CC BY)
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