Protein kinases are the largest class of enzymes in the human genome. These enzymes regulate almost all cellular processes by adding phosphate groups to proteins, thereby modifying the activity, localization, and overall function of their targets. Consequently, abnormal activity of kinases underlies many diseases including cancer.
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Proteome Profiler Human Phospho-Kinase Array Kit
R&D Systems | Catalog # ARY003C
Contains 4 sets of 2 membranes - each spotted in duplicate with antibodies against 37 different kinases and 2 related total proteins
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Key Product Details
Species
Human
Product Summary for Proteome Profiler Human Phospho-Kinase Array Kit
Kit Summary
A membrane-based antibody array for the parallel determination of the relative levels of human protein kinase phosphorylation. Validated for analyte detection in cell lysates.
Key Benefits
- Detects phosphorylation of 37 human kinases and total amounts of 2 related proteins simultaneously
- Requires no specialized equipment
Principle of the Assay
The Proteome Profiler Human Phospho-Kinase Array Kit is a membrane-based sandwich immunoassay. Capture antibodies spotted in duplicate on nitrocellulose membranes bind to specific target proteins present in the sample (Step 1). Captured proteins are detected with biotinylated detection antibodies (Step 2) and then visualized using chemiluminescent detection reagents (Step 3). The signal produced is proportional to the amount phosphorylation in the bound analyte.
Why Use an Antibody Array to Detect Receptor Phosphorylation?
Determining the phosphorylation of multiple kinases in a single sample can be expensive, time consuming and can require specialized equipment. Performing multiple immunoprecipitations and Western blots requires time, labor, and reagents. The use of a multiplex antibody array to detect multiple phosphorylations in a single sample can be cost-effective and also save time and sample.
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.
Stability and Storage
Store the unopened kit at 2°C to 8°C. Do not use past kit expiration date.
| Simultaneously detect the relative phosphorylation of these proteins in a single sample. | ||
|---|---|---|
| Akt 1/2/3 (S473) | HSP60 | PRAS40 (T246) |
| Akt 1/2/3 (T308) | JNK 1/2/3 (T183/Y185, T221/Y223) | Pyk2 (Y402) |
| beta-Catenin | Lck (Y394) | RSK1/2 (S221/S227) |
| Chk-2 (T68) | Lyn (Y397) | RSK1/2/3 (S380/S386/S377) |
| c-Jun (S63) | MSK1/2 (S376/S360) | Src (Y419) |
| CREB (S133) | p38 alpha (T180/Y182) | STAT1 (Y701) |
| EGF R (Y1086) | p53 (S15) | STAT2 (Y690) |
| eNOS (S1177) | p53 (S392) | STAT3 (S727) |
| ERK1/2 (T202/Y204, T185/Y187) | p53 (S46) | STAT3 (Y705) |
| Fgr (Y412) | P70 S6 Kinase (T389) | STAT5a/b (Y699) |
| GSK-3 alpha/beta (S21/S9) | p70 S6 Kinase (T421/S424) | STAT6 (Y641) |
| GSK-3 beta (S9) | PDGF R beta (Y751) | WNK-1 (T60) |
| HSP27 (S78/S82) | PLC gamma-1 (Y783) | Yes (Y426) |
Assays for analytes represented in the Human Phospho-Kinase Array Kit
| Analyte | DuoSet® IC ELISA Development Systems (Total) | DuoSet® IC ELISA Development Systems (Phospho) |
Quantikine® ELISA Kits |
|---|---|---|---|
| Akt (S473) | DYC887B | ||
| Akt (T308) | |||
| beta-Catenin | DYC1329 | ||
| Chk2 (T68) | DYC1626 | ||
| c-Jun (S63) | |||
| CREB (S133) | DYC2510 | ||
| EGF R (Y1086) | DYC1854 | DEGFR0 | |
| eNOS (S1177) | |||
| ERK1/2 (T202/Y204, T185/Y187) | DYC1018B | ||
| Fgr (Y412) | |||
| GSK-3 beta (S9) | |||
| GSK-3 alpha/beta (S21/S9) | DYC2157 | DYC2630 | |
| HSP27 (S78/S82) | DYC1580 | DYC2314 | |
| HSP60 | DYC1800 | ||
| JNK pan (T183/Y185, T221/Y223) | DYC1205 | DYC1387 | |
| Lck (Y394) | |||
| Lyn (Y397) | DYC3936 | ||
| MSK1/2 (S376/S360) | |||
| p38 alpha (T180/Y182) | DYC8691B | DYC869B | |
| p53 (S15) | DYC1043 | DYC1839 | |
| p53 (S46) | DYC1043 | DYC1489 | |
| p53 (S392) | DYC1043 | DYC2996 | |
| p70 S6 Kinase (T389) | DYC896 | ||
| p70 S6 Kinase (T421/S424) | DYC8962 | DYC8965 | |
| PDGF R beta (Y751) | DYC385 | DYC3096 | |
| PLC gamma-1 (Y783) | |||
| PRAS40 (T246) | DYC6890 | ||
| PYK2 (Y402) | |||
| RSK1/2 (S221/S227) | |||
| RSK1/2/3 (S380/S386/S377) | DYC889B | ||
| Src (Y419) | DYC2685 | ||
| STAT1 (Y701) | |||
| STAT2 (Y690) | |||
| STAT3 (S727) | |||
| STAT3 (Y705) | DYC4607B | ||
| STAT5a/b (Y694/Y699) | |||
| STAT6 (Y641) | |||
| WNK-1(T60) | DYC4720 | ||
| Yes (Y426) |
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Scientific Data Images for Proteome Profiler Human Phospho-Kinase Array Kit
Detection of Human Protein Kinase Phosphorylation in A549 Cell Line.
A549 human lung adenocarcinoma cells were either left untreated or exposed to 50 J/m2 of UV light followed by a 24 hour recovery before lysis.
Detection of Human Protein Kinase Phosphorylation in Daudi Cell Line.
Daudi human B cell lymphoma cells were either left untreated or treated with 500 U/ml Recombinant Human IFN-alpha 2a (R&D Systems, Catalog # 11100-1) for 5 minutes.
Detection of Human Protein Kinase Phosphorylation in Jurkat Cell Line.
Jurkat human acute T cell leukemia cells were either left untreated or treated with 100 ng/mL PMA and 0.5 mM Ionomycin for 20 minutes.
Detection of Human Protein Kinase Phosphorylation in MCF-7 Cell Line.
MCF-7 human breast cancer cells were either left untreated or treated with 100 ng/mL of recombinant human IGF-1 (R&D Systems®, Catalog # 291-G1) for 1 hour.
Formulation, Preparation, and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Kinase Assay Kits
Additional Kinase Assay Kits Products
Product Documents for Proteome Profiler Human Phospho-Kinase Array Kit
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Proteome Profiler Human Phospho-Kinase Array Kit
For research use only
Related Research Areas
Citations for Proteome Profiler Human Phospho-Kinase Array Kit
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Customer Reviews for Proteome Profiler Human Phospho-Kinase Array Kit (12)
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Verified Customer | Posted 12/05/2024
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Verified Customer | Posted 12/05/2024
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Verified Customer | Posted 04/24/2023
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Verified Customer | Posted 08/02/2022Used human lymphoma samples! Worked fine!
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Verified Customer | Posted 08/02/2022Used this kit with mouse lymphoma cell lines and it worked!
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Verified Customer | Posted 09/16/2021
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Verified Customer | Posted 01/27/2021
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Verified Customer | Posted 11/06/2020
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Verified Customer | Posted 03/18/2020Some of the antibodies lack sensitivity. Independent comprobation using the exact same protocol (lysis buffer, etc) and conventional WB rendered higher activation than the kit's
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Verified Customer | Posted 01/21/2020
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FAQs for Proteome Profiler Human Phospho-Kinase Array Kit
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A: ARY003C contains 8 membranes because it is divided into parts A and B with 4 membranes each. Both parts are spotted with different antibodies to avoid interference and cross reactivity as well as maximize sensitivity. Parts A and B each contain 39 different capture antibodies printed in duplicate and should be used together for optimal analysis efficiency. A total of four samples can be run with this kit.