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les1_ppt_blackwhite - Experimental Molecular Cell Biology...

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Experimental Molecular Cell Biology 2008-2009 Prof. Dr. R. Beyaert Dr. I. Carpentier
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When do the lectures take place… I. Carpentier I. Carpentier R. Beyaert I. Carpentier I. Carpentier I. Carpentier B. De Craene I. Carpentier R. Beyaert R. Beyaert R. Beyaert 1 2 Holiday 3 4 (14-15.15h aud 4) 5 6 No lecture 7 Easter holidays Easter holidays 8 9 10 11 9/2 16/2 23/2 2/3 9/3 16/3 23/3 30/3 6/4 13/4 20/4 27/4 4/5 11/5 Who? lesson date
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Work colleges Purposes: Problem solving + implementation of the theory by means of cases Gain insight in the theory Interpretation of article figures, data,… Articles on protein modifications: presentation + discussion Obliged attendance When + where: 10/3 and 21/4 from 14-18h 4th floor (or Ada Lovelace PC room)
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Evaluation Theory: closed book 19/20 • Work college 1/20
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INTRODUCTION Imagine…..
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Cases Case 1: Your supervisor has heard at a congress about this very interesting promising new gene. He wants you to write a project on the protein and start to perform experiments. After a literature search you only know that the encoded protein upon overexpression is involved in the activation of a certain transcription factor….
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Case 2: Suppose a biotech company recruits you. Your job is to develop and optimize high through put cell- based assays to screen drug databases for drugs that inhibit inflammation…..
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Questions: How to start? • Logical steps? • Techniques? Design of experiments? • ……
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Conclusion: Profound knowledge of molecular biomedical techniques are indispensable!
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What EMCB course can mean for you: Insight in advanced molecular and cell biological techniques commonly used in academic and industrial research Sufficient background information to be able to design experiments Better understanding and more critical evaluation of scientific literature
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EMCB course will discuss…. Molecular biomedical techniques: – theories and principles – Applications – Practical aspects and limitations
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What about case 1 … 1. Scientific literature search on Pubmed 2. Bio-informatics 3. Setup of experiment
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Experiment setup: some guidelines Optimize, confirm, alternatives •Choose model organism, tissue, cell type •Choose the right conditions ( e.g. stimulus, time kinetics, transfection protocols, ...) •Choose the “best” technique (SWOT) •+/- controls •Perform exp in Tissue Culture conform GCCP •Leave intact, fix or lyse cells: techniques
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Biological source material Living animals Generation of extract Protein RNA DNA 1 9,10,11 2 7,8 Protein 4,5,6 3 Cell death and proliferation techniques phosphorylation ubiquitination sumoylation acetylation methylation lipid modification ADP ribosylation Reporter systems and detection methods Epitope tagging, physical and gene-based protein-protein interaction methods: - Physical (affinity chromatography, TAP, pull down, coIP, affinity blotting, crosslinking, SPA, Biacore, protein arrays) -Gene-based (protein probing,
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  • Spring '10
  • JohanGrooten
  • Cell culture, laminar flow hoods, I. Carpentier I. Carpentier R. Beyaert I. Carpentier I. Carpentier I. Carpentier B. De Craene

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