Protocols and Applications Guide (US Letter Size) - Promega

||||||| 7Cell Signaling
CONTENTS
I. Introduction 1
A. The MAPK Pathways 1
B. The Phosphoinositol 3-Kinases (PI3-Ks) 1
C. Investigating Phosphatases and Kinases as Potential
Therapeutic Targets 3
II. Kinase Activity Assays 3
A. Luminescent Kinase Assays 3
B. Fluorescent Kinase Assays 6
III. Radioactive Kinase Assays 8
A. SAM2® Biotin Capture Membrane and Biotin Capture
Plate 8
B. SignaTECT® Protein Kinase Assay Systems 9
C. Other Kinase Assay Formats (non-radioactive) 10
IV. Phosphorylation-Specific Antibodies 11
A. Phosphorylation-Specific Antibodies in MAPK Signaling
Pathways 11
B. Phosphorylation-Specfic CaM KII Antibody 13
V. Kinase Inhibitors 15
A. MEK Inhibitor U0126 15
B. PD 98059 15
C. SB 203580 15
D. PI3 Kinase Inhibitor LY 294002 16
E. cAMP-Dependent Protein Kinase (PKA) Peptide
Inhibitor 16
F. InCELLect® AKAP St-Ht31 Inhibitor Peptide 16
G. Myristoylated Protein Kinase C Peptide Inhibitor 16
H. Olomoucine cdc2 Protein Kinase Inhibitor 17
VI. Phosphatase Assays 17
A. Fluorescent Phosphatase Assays 17
B. Colorimetric Phosphatase Assays 18
VII. References 19
Protocols & Applications Guide
www.promega.com
rev. 3/09
PROTOCOLS & APPLICATIONS GUIDE