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.
Proteome Profiler Human Phospho-Kinase Array Kit
R&D Systems | Catalog # ARY003B
Discontinued Product
ARY003B has been discontinued.
An alternative/replacement product is available:
ARY003C. View all Kinase Assay Kits products.
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 43 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). Phosphorylation of captured proteins is detected with biotinylated phospho-specific 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) | Hck (Y411) | PLC gamma-1 (Y783) |
| Akt 1/2/3 (T308) | HSP27 (S78/S82) | PRAS40 (T246) |
| AMPK alpha1 (T183) | HSP60 | Pyk2 (Y402) |
| AMPK alpha2 (T172) | JNK 1/2/3 (T183/Y185, T221/Y223) | RSK1/2/3 (S380) |
| beta-Catenin | Lck (Y394) | Src (Y419) |
| Chk-2 (T68) | Lyn (Y397) | STAT2 (Y689) |
| c-Jun (S63) | MSK1/2 (S376/S360) | STAT3 (S727) |
| CREB (S133) | p27 (T198) | STAT3 (Y705) |
| EGF R (Y1086) | p38 alpha (T180/Y182) | STAT5a (Y699) |
| eNOS (S1177) | p53 (S15) | STAT5a/b (Y699) |
| ERK1/2 (T202/Y204, T185/Y187) | p53 (S392) | STAT5b (Y699) |
| FAK (Y397) | p53 (S46) | STAT6 (Y641) |
| Fgr (Y412) | P70 S6 Kinase (T389) | TOR (S2448) |
| Fyn (Y420) | p70 S6 Kinase (T421/S424) | WNK-1 (T60) |
| GSK-3 alpha/beta (S21/S9) | PDGF R beta (Y751) | 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) | |||
| AMPK alpha1 (T183) | DYC3197 | DYC3528 | |
| AMPK alpha2 (T172) | |||
| beta-Catenin | DYC1329 | DYC1329 | |
| Chk2 (T68) | DYC1626 | ||
| c-Jun (S63) | |||
| CREB (S133) | DYC2510 | ||
| EGF R (Y1086) | DYC1854 | DEGFR0 | |
| eNOS (S1177) | |||
| ERK1/2 (T202/Y204, T185/Y187) | DYC1018B | ||
| FAK (Y397) | DYC4467 | ||
| Fgr (Y412) | |||
| Fyn (Y420) | |||
| GSK-3 alpha/beta (S21/S9) | DYC2157 | DYC2630 | |
| Hck (Y411) | |||
| HSP27 (S78/S82) | DYC1580 | DYC2314 | |
| HSP60 | DYC1800 | ||
| JNK pan (T183/Y185, T221/Y223) | DYC1205 | DYC1387 | |
| Lck (Y394) | |||
| Lyn (Y397) | DYC3936 | ||
| MSK1/2 (S376/S360) | |||
| p27 (T198) | DYC2256 | ||
| 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) | |||
| RSK1/2/3 (S380/S386/S377) | DYC889B | ||
| Src (Y419) | DYC2685 | ||
| STAT2 (Y689) | |||
| STAT3 (S727) | |||
| STAT3 (Y705) | DYC4607B | ||
| STAT5a (Y694) | |||
| STAT5a/b (Y694/Y699) | |||
| STAT5b (Y699) | |||
| STAT6 (Y641) | |||
| TOR (S2448) | DYC1665 | ||
| WNK-1(T60) | DYC4720 | ||
| Yes (Y426) | DYC3929 |
Data Examples
 View Larger Image |
Figure 1. MCF-7 human breast cancer cells were either left untreated or exposed to 50 J/m2 of UV light followed by a 4 hour recovery before lysis. |
 View Larger Image |
Figure 2. A431 human epithelial carcinoma cells were either left untreated or treated with 200 ng/mL recombinant human (rh) EGF (Catalog # 236-EG) for 5 minutes. |
 View Larger Image |
Figure 3. MCF-7 human breast cancer cells were either left untreated or treated with 100 ng/mL of rhIGF-I (Catalog # 291-G1) for 1 hour. |
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Formulation, Preparation, and Storage
Shipping
The product is shipped with polar packs. 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 (23)
4.8 out of 5
23 Customer Ratings
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Verified Customer | Posted 10/16/2020
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Verified Customer | Posted 08/08/2019
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Verified Customer | Posted 06/26/2019
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Verified Customer | Posted 06/05/2019
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Verified Customer | Posted 06/02/2019Prostae cancer cell lines were used and 200 ug lysate for each samples per membrane was applied.
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Verified Customer | Posted 04/11/20192xce arrays purchased circa Jan 2018
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Verified Customer | Posted 04/05/2019
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Verified Customer | Posted 03/05/2019
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Verified Customer | Posted 01/31/2019
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Verified Customer | Posted 11/29/2018
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Verified Customer | Posted 09/13/2018Great tool for evaluating cell signaling changes. We used this kit with LS174T cells. Kit protocol was easy to follow, and after imaging the resulting spots were very clear and easy to analyze. Phosphorylation of one of the identified signaling proteins was verified using an independent western blot. I would recommend this product
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Verified Customer | Posted 07/18/2018
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Verified Customer | Posted 04/18/2018
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Verified Customer | Posted 04/05/2018
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Verified Customer | Posted 03/22/2018
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Verified Customer | Posted 10/20/2017
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Verified Customer | Posted 02/13/2017Great kit and ok value for money considering the amount of data you get!
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Verified Customer | Posted 02/12/2017Used with human samples. Image shown was at 1 minute. Other 'dots' appeared at increased exposure. Kit very good overall. Good quality result and easy to use
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Verified Customer | Posted 01/03/2017This array is an excellent tool for screening potential phospho-kinases and related targets. However, it would be nice that if the total level of non-phosphorylated proteins be visualized at the same time on the membrane. In this case, the phosphorylated signals can be normalized.
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Verified Customer | Posted 10/28/2016Antibodies could not be sourced to validate some of the targets.
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Verified Customer | Posted 10/14/2016
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Verified Customer | Posted 08/09/2016
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Sample Tested: Cell LysatesVerified Customer | Posted 10/26/2015This was control and didnt show any non specific bindings. <br />Specificity: Specific<br />Sensitivity: Sensitive<br />Buffer: BSA<br />Dilution: 1:1000
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