hitachiorigenewebinar6-28-10-ver3-100701181149-phpapp02.pptVIP

* * * * * * * * * * * * * * * * * * * * * * * * * * * * * * TATA Pol II TBP (TATA Binding Protein) TFIID Eukaryotic Promoter Exons mRNA TATA Pol II TBP (TATA Binding Protein) TFIID Eukaryotic Promoter Exons mRNA Low Expression Levels High Expression Levels Transcription Factors Bind to DNA to Regulate Transcription Gel shift assays (EMSAs) and Western blots gel based, not multiplexed, large sample requirement Reporter gene assays requires cell culture, not multiplexed, delayed kinetics ELISAs quantitative and automatable, but not easily multiplexed Traditional Methods for Measuring TF Activity Are Inadequate Transcription Factor Profiling Assays 0 200 400 600 800 1000 1200 NF-kb CREB ISRE EGR ETS alt AP4 NF-1 Octamer GATA alt PPAR AP1 SMAD 1 TATA SMAD 3 STAT NBS1 MTB1b HNF-4 AML-1 FREAK SAF CRE ATF1 TAL1 E2F1-5 E2F6 MYCX NFY1 ER Assay Net MFI StRE Transcription Factor Profile Obtained with Recombinant NF-kB p50 and NF-1 Slope = 1.07 Lysate Analysis C U T PPAR EMSA C U T HL-60 Cells treated with PMA/IM Multiplexed Immunoassays Multiplex Measurement of Protein Concentration in Biological Fluids Antibody Sandwich Assay Luminex Immunoassays Produced from TrueORF cDNA clones 5,000 full length human proteins Expressed in HEK293 cells Optimal preservation of protein structure, post-translational modifications and functions Full Length Human Proteins Dedicated mAb center Capacity: 200 immunizations per month Authentic proteins as antigens Full length proteins made in human cells Extensive validation WB: 9 different cell lines IHC: 24 different tissues IF Flow Luminex Goal: produce high quality mAb to every human proteins in 5-10 years mAb for Every Human Protein Analyte Name Symbol Platelet-derived growth factor AB PDGFAB Platelet-derived growth factor BB PDGFBB Fibroblast growth factor 4 FGF4 Vascular endothelial growth factor D VEGFD Fibroblast growth factor 2 FGF2 Epidermal growth factor EGF Hepatocyte growth factor HGF Flt-3 ligand FLT3LG Ang