Discuss, giving specific examples, the use of antibodies to study epidermal growth factor receptor signalling

Order Description exam (try the structure of essay to be easy to memorise) not more than 800 words some the references should be updated within 3-4 years and tray to avoid review articles Discuss, giving specific examples, the use of antibodies for the study of cell signalling involving protein kinase cascades. Your answer should include any advantages and/or disadvantages and reference to practical considerations when using such antibodies. General introduction to the role of protein kinases in signal transduction including the two main families (serine/threonine kinases and tyrosine kinases). Reference to the general uses of antibodies to study cell signaling pathways e.g. the use of antibodies to immunoprecipitate cell signalling proteins and the localisation of proteins via immunohistochemistry, immunocytochemistry may also be included. The main focus of the answer should highlight the importance of protein phosphorylation in cell signaling lead to the development of anti phospho-tyrosine antibodies (attempts to produce anti phospho-serine and anti phospho-threonine – less successful). Also the development of phospho- specific antibodies has revolutionized the study of signaling pathways using antibodies. Advantages: no need to use isotopes, can target specific phosphorylation sites. Disadvantages: may miss activation of a specific protein kinase if additional (and perhaps unknown) pathways of activation exist that are independent of the phospho-specific antibody used. Practical considerations i.e. must inhibit phosphatase activity, choice of immunoprecipitating antibody, protein levels etc. Specific examples include the MAPK pathway and BAD phosphorylation. Additional marks awarded for evidence of background reading.