Development of Fluorescence Polarization and FRET Assays for Tyrosine and Serine/Threonine Kinases

Dr. Jinzi J. Wu
Novartis Institute for Biomedical Research
Summit, New Jersey

4/99

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Table of Contents

Development of Fluorescence Polarization and FRET Assays for Tyrosine and Serine/Threonine Kinases
Dr. Jinzi J. Wu
Novartis Institute for Biomedical Research
Summit, New Jersey

FP and FRET: Assay diagrams

FRET for TK1: Assay optimization

Filtration, SPA and FRET assays for TK1

FRET for TK1: Reagent profiling

Summary of the FRET assay of TK1

FP for TK2: Sensitivity of the LJL Analyst

FP, FRET and SPA for TK2: Enzyme titration

IC50’s in FP, FRET and SPA assays

Summary of FP and FRET assays for TK2

Development of FP assays for S/T kinases

FP for PKC: Enzyme titration

FP for PKC: Inhibition of kinase activity

FP for PKC: Conclusions

Recognition motifs for S/T kinases

FP for Akt: Tracer and antibody titration

FP for Akt: Specificity of antibody

FP for Akt: Substrate and enzyme titration

FP and SPA for Akt: Inhibition curves

Summary of FP assay for Akt

Acknowledgments

Author: Jinzi Wu

Other information:
LRIG Mid Atlantic April 99 Meeting

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