Explain the rationale for using the primary and secondary antibodies in the IHC method as described by Soysal et al (2013). Illustrate the interactions using schematic diagrams.

Questions:

Q1) EpCam (Epithelial cell adhesion molecule) has been associated with several cancers including breast cancer. In the paper by Soysal et al (2013) entitled ‘EpCAM expression varies significantly and is differentially associated with prognosis in the luminal B HER2⁺, basal-like, and HER2 intrinsic subtypes of breast cancer‘ the authors investigated levels of EpCam using immunohistochemistry.

Read the paper and method section very carefully (linked to paper:

http://www.nature.com/bjc/journal/v108/n7/pdf/bjc201380a.pdf) and answer the following questions:

Insert your answers in the spaces below, adding more lines as required.

(a) Explain the rationale for using the primary and secondary antibodies in the IHC method as described by Soysal et al (2013). Illustrate the interactions using schematic diagrams.

(5 marks)

Your answer

(b) Identify and explain the rationale underlying at least three main controls and/or optimisation procedures which are required to reduce false positive and/or false negative results.

(10 marks)

Complete the table

(c) What is the scientific basis of using H & E staining in immunohistochemistry                                                                                                                (5 marks)