Introduction to IHC antibody validation for Cancer biomarkers

Immunohistochemistry determines the cellular or subcellular localization of a specific protein. This technique plays a vital role in diagnostics and is increasingly used to target drug therapies as a quick and cost-effective assay. Biomarkers for conditions such as cancer are commonly analyzed via IHC to yield essential diagnostic and prognostic information and to monitor treatments.

Interpreting positive and negative signals in IHC assays can be difficult. In addition to the acknowledged problems of antigen retrieval in paraffin-embedded sections, the differential availability of antigens between different assay formats means that an antibody recognizing a single band on a western blot may recognize multiple proteins in IHC.

There is no easy way around these problems. The usual approach is to compare samples to both positive and negative control sample staining and to demonstrate that similar signals can be obtained using multiple antibodies against the same target.

A crucial step prior to IHC analysis is antibody validation; this will ensure your antibody is specific to the cancer biomarker of interest and sufficiently sensitive to allow IHC analysis over the required dynamic range demanded by the pathology.

Early validation of your cancer biomarker antibody offers confidence in your results and allows a better understanding of the target. Work produced with validated antibodies ensures long-term reproducibility of results and is quicker to transition to the clinical setting.

At Abcam, we perform rigorous antibody validation for IHC to guarantee antibody specificity, sensitivity and reproducibility. We include numerous reviews direct from researchers highlighting how our antibodies perform hands-on across many different labs.

References

1. Yu, J.,, Kane, S.,, et al. Mutation-specific antibodies for the detection of EGFR mutations in non-small-cell lung cancer Clin. Cancer Res. 15 ,3023-3028 (2009)
2. Howat, W. J.,, Lewis, A.,, et al. Antibody validation of immunohistochemistry for biomarker discovery: Recommendations of a consortium of academic and pharmaceutical based histopathology researchers. Methods 70 (1),34-38 (2014)
3. Deutsch, E. W.,, Ball, C. A.,, et al. Minimum information specification for in situ hybridization and immunohistochemistry experiments (MISFISHIE). Nat. Biotechnol.  26 ,305-312 (2008)