CST Guide:

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Table of ContentS
I: Research Areas
01 Gene Expression, Epigenetics, and Nuclear Function 20
Chromatin/Epigenetic Regulation 20
Commonly Studied Chromatin/Epigenetic Regulation Targets 24
Protein Acetylation Pathway 26
Histone Lysine Methylation Pathway 27
Histone Modifications Table 28
Examples of Crosstalk Between Post-Translational Modifications 31
Pathway
Translational Control 32
Commonly Studied Translational Control Targets 35
Translational Control: Overview Pathway 36
Translational Control: Regulation of elF4E and p70 S6K Pathway 37
Translational Control: Regulation of elF Pathway 38
Nuclear Receptors 39
Commonly Studied Nuclear Receptor Targets 39
Nuclear Receptors Signaling Pathway 41
02 Signaling 42
MAP Kinase Signaling 42
Commonly Studied MAP Kinase Targets 45
MAPK/Erk in Growth and Differentiation Pathway 44
Signaling Pathways Activating p38 MAP Kinase Pathway 47
SAPK/JNK Signaling Pathway 48
PI3 Kinase/Akt Signaling 49
Commonly Studied Akt Targets 51
PI3 Kinase/Akt Signaling Pathway 53
mTOR Signaling Pathway 54
Akt Substrates Pathway and Table 55
Additional Binding Partners Table 65
Tyrosine Kinases and Associated Phosphatases 66
Commonly Studied Tyrosine Kinases Targets 69
Tyrosine Kinases Kinase-Disease Associations Table 71
ErbB/HER Signaling Pathway 74
G Protein-Coupled Receptors 75
Commonly Studied GPCR Targets 77
G Protein-Coupled Receptor Signaling: Overview Pathway 78
G Protein-Coupled Receptor Signaling to MAP Kinase/Erk Pathway 79
Calcium, cAMP, and Lipid Signaling 80
Commonly Studied Calcium, cAMP, and Lipid Signaling Targets 83
Calcium, cAMP, and Lipid Signaling Kinase-Disease Associations 84
Table
03 Cell Growth and Death 86
Apoptosis 86
Commonly Studied Apoptosis Targets 88
Regulation of Apoptosis Overview Pathway 90
Inhibition of Apoptosis Pathway 91
Death Receptor Signaling Pathway 92
Mitochondrial Control of Apoptosis Pathway 93
Autophagy 94
Commonly Studied Autophagy Targets 96
Autophagy Signaling Pathway 97
Cell Cycle, Checkpoint Control, and DNA Damage 98
Commonly Studied Cell Cycle Targets 100
Cell Cycle/Checkpoint Control Kinase-Disease Associations 102
Cell Cycle Control: G1/S Checkpoint Pathway 104
Cell Cycle Control: G2/M DNA Damage Checkpoint Pathway 105
04 Cell Biology 106
Adhesion and Extracellular Matrix 106
Commonly Studied Adhesion Targets 109
Adherens Junction Dynamics Pathway 110
Cytoskeletal Regulation 111
Commonly Studied Cytoskeletal Regulation Targets 113
Regulation of Actin Dynamics Pathway 115
Regulation of Microtubule Dynamics Pathway 116
Protein Folding and VesIcle Trafficking 117
Commonly Studied Protein Folding and Vesicle Trafficking Targets 119
Ubiquitin and Ubiquitin-like Proteins 120
Commonly Studied Ubiquitin Targets 123
Ubiquitin/Proteasome Pathway 124
Ubiquitin Ligase Table 125
Deubiquitinase Table 135
05 Cellular Metabolism 140
Commonly Studied Cellular Metabolism Targets 142
Insulin Receptor Signaling Pathway 144
Warburg Effect Pathway 145
AMPK Signaling Pathway 146
AMPK Substrates Table 147
06 Development and Differentiation 152
Commonly Studied Development and Differentiation Targets 158
Wnt/β-Catenin Signaling Pathway 160
Notch Signaling Pathway 161
Hippo Signaling Pathway 162
Hedgehog Signaling Pathway 163
TGF-β Signaling Pathway 164
ESC Pluripotency Differentiation Pathway 165
Angiogenesis 166
Commonly Studied Angiogenesis Targets 170
Angiogenesis Signaling in Tumor Neovascularization Pathway 171
07 Immunology and Inflammation 172
Commonly Studied Immunology and Inflammation Targets 174
Jak and Cytokine Receptor Mutants Table 176
Jak/Stat Utilization Table 177
B Cell Receptor Signaling Pathway 178
T Cell Receptor Signaling Pathway 179
Toll-like Receptor Signaling Pathway 180
Jak/Stat Signaling: IL-6 Receptor Family Pathway 181
NF-κB Signaling Pathway 182
Tumor Immunology Pathway 183
08 Neuroscience 184
Commonly Studied Neuroscience Targets 187
Vesicle Trafficking in Presynaptic Neurons: Synchronous Release 189
Pathway
Amyloid Plaque and Neurofibrillary Tangle Formation in Alzheimer’s 190
Disease Pathway
Dopamine Signaling in Parkinson’s Disease Pathway 191
II: Antibody Applications
09 Chromatin Immunoprecipitation (ChIP) 194
ChIP General Protocol 196
ChIP Troubleshooting Guide 200
10 Flow Cytometry (F) 202
Flow Cytometry General Protocol 204
Flow Cytometry Alternate Protocol (for combined staining
204
of intracellular proteins and cell surface markers in blood)
11 Immunofluorescence (IF) 206
Cultured Cells (Immunocytochemistry, IF-IC) Protocol 210
Frozen/Cryostat Tissue Sections (IF-F) Protocol 211
Paraffin Tissue Sections (IF-P) Protocol 212
In-Cell Western Protocol 213
12 Immunohistochemistry (IHC) 214
IHC Paraffin Protocol (using SignalStain ® Boost Detection Reagent) 216
IHC Frozen Protocol (using SignalStain ® Boost Detection Reagent) 217
IHC Troubleshooting Guide 219
III: Workflow Tools
17 Exploration 250
Using PhosphoSitePlus ® online database
18 Investigation 254
Using motif and post-translational modification-specific antibodies
19 Discovery 258
Using peptide immunoaffinity enrichment and LC-MS/MS to study
post-translational modifications (PTMs)
20 Identification 262
Using Chromatin IP (ChIP) to identify protein-DNA interactions
21 Profiling 264
Using antibody arrays
22 Modulation 266
Using chemical activators and inhibitors, cytokines, and growth factors
IV: Additional Information
28 About CST 282
Behind the Antibodies 282
Corporate Social Responsibility 282
Sustainability 283
Environmental Awareness and Support 283
29 CST Nature Conservancy 284
Conservancy Efforts & Resources 284
30 Publications by CST Scientists 290
31 Index 294
Pathway Diagrams and Tables 294
Protocols and Troubleshooting Guides 294
Targets 295
Trademarks, Terms and Conditions 301
13 Sandwich ELISA 220
PathScan ® Sandwich ELISA Colorimetric Protocol 221
PathScan ® Sandwich ELISA Chemiluminescent Protocol 223
PathScan ® Sandwich ELISA Antibody Pair Protocol 224
14 Western Blotting (WB) 226
WB General Protocol 228
WB Fluorescent Protocol 230
WB Troubleshooting Guide 232
15 Immunoprecipitation (IP) 234
IP Native Protein Protocol 235
IP Denatured Protein Protocol 237
16 Control Treatments and Cell Lines 238
23 Localization & Classification 269
Using conjugated primary antibodies
24 Screening & Quantification 272
Using ELISA to screen and quantify cellular responses at the target level
25 Monitoring 274
Using cell assays to monitor the health of cells and their response to
multiple challenges
26 Verification 277
Using siRNA-mediated knockdown to verify function
27 Customization 278
Antibodies in a carrier-free formulation for specific assay platforms
32 Diagram and Table Keys 302
Pathway Diagram Key 302
Application Key 303
Reactivity Key 303
Amino Acid Properties Table 303
33 Contact Information 304
Global Headquarters 304
Ordering and Customer Service 304
Technical Support 304
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