Proteome Profiler Human Phospho-Kinase Array Kit

Contains 4 sets of 2 membranes - each spotted in duplicate with antibodies against 37 different kinases and 2 related total proteins
Catalog # Availability Size / Price Qty
ARY003C
Detection of human protein kinase phosphorylation in A549 Cell Line.
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Citations (389)
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Proteome Profiler Human Phospho-Kinase Array Kit Summary

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.

Troubleshooting Guide

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)   DYC3929  

Specifications

Source
N/A
Shipping Conditions
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.
Species
Human

Product Datasheets

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Scientific Data

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.

Citations for Proteome Profiler Human Phospho-Kinase Array Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

389 Citations: Showing 1 - 10
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  18. PTPN2 copper-sensing rapidly relays copper level fluctuations into EGFR/CREB activation and associated CTR1 transcriptional repression
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  36. Oxidized Proteins Differentially Affect Maturation and Activation of Human Monocyte-Derived Cells
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  37. BCG vaccinations drive epigenetic changes to the human T cell receptor: Restored expression in type 1 diabetes
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    Science Advances, 2022-11-16;8(46):eabq7240.  2022-11-16
  38. Colon tumour cell death causes mTOR dependence by paracrine P2X4 stimulation
    Authors: M Schmitt, F Ceteci, J Gupta, M Pesic, TW Böttger, AM Nicolas, KB Kennel, E Engel, M Schewe, A Callak Kir, V Petrocelli, Y Dabiri, J Varga, M Ramakrishn, M Karimova, A Ablasser, T Sato, MC Arkan, FJ de Sauvage, FR Greten
    Nature, 2022-11-16;0(0):.  2022-11-16
  39. Extracellular Vesicles Isolated from Malignant Mesothelioma Cancer-Associated Fibroblasts Induce Pro-Oncogenic Changes in Healthy Mesothelial Cells
    Authors: T Chernova, S Grosso, XM Sun, AR Tenor, JZ Cabeza, A Craxton, EL Self, A Nakas, K Cain, M MacFarlane, AE Willis
    International Journal of Molecular Sciences, 2022-10-18;23(20):.  2022-10-18
  40. Loss of NECTIN1 triggers melanoma dissemination upon local IGF1 depletion
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    Nature Genetics, 2022-10-13;0(0):.  2022-10-13
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  43. Post-translational dysregulation of glucose uptake during exhaustive cycling exercise in vastus lateralis muscle of healthy homozygous carriers of the ACE deletion allele
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    The Journal of Biological Chemistry, 2022-08-07;298(9):102353.  2022-08-07
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  50. Tsp2 Facilitates Tumor-associated Fibroblasts Formation and Promotes Tumor Progression in Retroperitoneal Liposarcoma
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    International journal of biological sciences, 2022-08-01;18(13):5038-5055.  2022-08-01
  51. The human amniotic epithelium confers a bias to differentiate toward the neuroectoderm lineage in human embryonic stem cells
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  52. Kayadiol exerted anticancer effects through p53-mediated ferroptosis in NKTCL cells
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    British Journal of Cancer, 2022-06-24;0(0):.  2022-06-24
  54. Peroxiredoxin IV plays a critical role in cancer cell growth and radioresistance through the activation of the Akt/GSK3 signaling pathways
    Authors: N Ding, H Jiang, P Thapa, Y Hao, A Alshahrani, D Allison, T Izumi, VM Rangnekar, X Liu, Q Wei
    The Journal of Biological Chemistry, 2022-06-10;0(0):102123.  2022-06-10
  55. GSK3beta palmitoylation mediated by ZDHHC4 promotes tumorigenicity of glioblastoma stem cells in temozolomide-resistant glioblastoma through the EZH2-STAT3 axis
    Authors: C Zhao, H Yu, X Fan, W Niu, J Fan, S Sun, M Gong, B Zhao, Z Fang, X Chen
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  56. Idelalisib activates AKT via increased recruitment of PI3Kdelta/PI3Kbeta to BCR signalosome while reducing PDK1 in post-therapy CLL cells
    Authors: MK Mamidi, H Mahmud, GP Maiti, MT Mendez, SM Fernandes, SK Vesely, J Holter-Cha, JR Brown, AK Ghosh
    Leukemia, 2022-05-14;0(0):.  2022-05-14
  57. m6A methylation mediates LHPP acetylation as a tumour aerobic glycolysis suppressor to improve the prognosis of gastric cancer
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    Cell Death & Disease, 2022-05-14;13(5):463.  2022-05-14
  58. Ephrin B Activate Src Family Kinases in Fibroblasts Inducing Stromal Remodeling in Prostate Cancer
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    Cancers, 2022-05-09;14(9):.  2022-05-09
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    Proteomics, 2011-05-18;11(12):2406-22.  2011-05-18
  368. IGF1R Signaling in Ewing Sarcoma Is Shaped by Clathrin-/Caveolin-Dependent Endocytosis.
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  369. Influence of leucine on protein metabolism, phosphokinase expression, and cell proliferation in human duodenum1,3.
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    Am. J. Clin. Nutr., 2011-04-20;93(6):1255-62.  2011-04-20
  370. Sulfiredoxin-Peroxiredoxin IV axis promotes human lung cancer progression through modulation of specific phosphokinase signaling.
    Authors: Wei Q, Jiang H, Xiao Z
    Proc. Natl. Acad. Sci. U.S.A., 2011-04-12;108(17):7004-9.  2011-04-12
  371. BH3 mimetics activate multiple pro-autophagic pathways.
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FAQs

  1. Why are there 8 membranes provided in ARY003C?

    • 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. 

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