Application Note

Immunohistochemical Detection of Phospho-HER-3 in Human Breast Cancer

Establishing reliable protocols for immunohistochemistry requires a defined evaluation of antibody specificity

Immunohistochemistry (IHC) is a valuable tool to analyze molecular processes in healthy and diseased tissues, especially in tumor tissue. However, the implementation of IHC entails potentially serious pitfalls, such as artificial molecular alterations in the tissue. To prevent misleading outcomes, Indivumed Services implements a standardized procedure to establish high-quality staining protocols, including a thorough selection and evaluation of antibodies using different approaches.

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• The precise selection of critical antibodies and attributes such as immunogenicity, purification status, and clonality for the predicted target specificity of antibodies.
  
  
• The useful tools of immunoblotting to determine cross-reactivity of antibodies with other proteins.
  
  
• Matched antibody reactivity with protein bands of the predicted molecular weight indicates target specificity. BT474 and SKBR3 cells have been selected as positive controls for immunoblotting of phospho-HER-3.

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