Introduction
Analyzing the phosphorylation profiles of kinases and their protein substrates is essential for understanding how cells recognize and respond to changes in their environment. The Human Phospho-Kinase Array (Catalog # ARY003B) is a rapid, sensitive, and economical tool to simultaneously detect the relative site-specific phosphorylation of 43 kinases and 2 related total proteins without performing numerous immunoprecipitations and Western blots. Capture antibodies were carefully selected using cellular extracts prepared from cell lines known to express the target protein. Each capture antibody is spotted in duplicate on a nitrocellulose membrane and the levels of phosphorylated protein are assessed using phospho-specific antibodies and chemiluminescent detection.
- Product Insert (PDF, 1.1MB)
- Troubleshooting Guide
- Assays for analytes represented in the Human Phospho-Kinase Array Kit
- Western blotting antibodies for analytes related to the Human Phospho-Kinase Array Kit
| Simultaneously detect the relative phosphorylation of these proteins in a single sample. | ||
| Akt (S473) | Hck (Y411) | PRAS40 (T246) |
| Akt (T308) | HSP27 (S78/S82) | Pyk2 (Y402) |
| AMPK alpha1 (T174) | HSP60 | RSK1/2/3 (S380/S386/S377) |
| AMPK alpha2 (T172) | JNK pan (T183/Y185, T221/Y223) | Src (Y419) |
| beta-Catenin | Lck (Y394) | STAT2 (Y689) |
| Chk-2 (T68) | Lyn (Y397) | STAT3 (S727) |
| c-Jun (S63) | MSK1/2 (S376/S360) | STAT3 (Y705) |
| CREB (S133) | p27 (T198) | STAT5a (Y694) |
| EGF R (Y1086) | p38 alpha (T180/Y182) | STAT5a/b (Y694/Y699) |
| eNOS (S1177) | p53 (S15) | STAT5b (Y699) |
| ERK1/2 (T202/Y204, T185/Y187) | p53 (S392) | STAT6 (Y641) |
| FAK (Y397) | p53 (S46) | TOR (S2448) |
| Fgr (Y412) | p70 S6 Kinase (T421/S424) | WNK-1 (T60) |
| Fyn (Y420) | PDGF R beta (Y751) | Yes (Y426) |
| GSK-3 alpha/beta (S21/S9) | PLC gamma-1 (Y783) | |
General Assay Principle
Carefully selected capture antibodies have been spotted in duplicate on nitrocellulose membranes. Cell lysate samples are diluted and mixed with a cocktail of biotinylated detection antibodies. The sample/antibody mixture is then incubated with the array. Any protein/detection antibody complex present is bound by its cognate immobilized capture antibody on the membrane. Streptavidin-Horseradish Peroxidase and chemiluminescent detection reagents are added, and a signal is produced in proportion to the amount of cytokine bound. Chemiluminescence is detected in the same manner as a Western blot.
Kit Contents
Each Human Phospho-Kinase Antibody Array Kit contains most of the necessary components for a ready-to-run assay without the need for specialized equipment.
- 8 Array Membranes (4 of membrane A and 4 of membrane B)
- 8-Well Multi-dish
- Lysis Buffer
- Array Buffers
- Wash Buffer
- Detection Antibody Cocktail A
- Detection Antibody Cocktail B
- Streptavidin-HRP
- Chemiluminescent Detection Reagents
- Transparency Overlay Template
- Detailed product datasheet with protocol
For a complete list of the kit contents and necessary materials, please see the Materials Provided/Other Supplies Required sections of the product datasheet.