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
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ARY003C
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Detection of human protein kinase phosphorylation in A549 Cell Line.
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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.

392 Citations: Showing 1 - 10
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  1. New metal complexes of 1H-benzimidazole-2-yl hydrazones: cytostatic, proapoptotic and modulatory activity on kinase signaling pathways
    Authors: Argirova, M;Cherneva, E;Mihaylova, R;Momekov, G;Yancheva, D;
    Archives of biochemistry and biophysics  2024-11-29
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    Research square  2024-10-15
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    bioRxiv : the preprint server for biology  2024-08-28
  5. PTPN2 copper-sensing relays copper level fluctuations into EGFR/CREB activation and associated CTR1 transcriptional repression
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    Nature communications  2024-08-13
  6. Comprehensive Analysis of Kisspeptin Signaling: Effects on Cellular Dynamics in Cervical Cancer
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  9. Selective targeting of IRAK1 attenuates low molecular weight hyaluronic acid-induced stemness and non-canonical STAT3 activation in epithelial ovarian cancer
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  12. Brain-targeted delivery of neuroprotective survival gene minimizing hematopoietic cell contamination: implications for Parkinson's disease treatment
    Authors: Lee, MH;Kang, S;Um, KH;Lee, SW;Hwang, H;Baek, K;Choi, JW;
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  14. GRB7 plays a promoting role in the progression of gastric cancer
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    Authors: Gonzalez Suarez, N;Fernandez-Marrero, Y;Hébert, MPA;Roy, ME;Boudreau, LH;Annabi, B;
    Cancer cell international  2023-10-13
  17. Fumarate hydratase functions as a tumor suppressor in endometrial cancer by inactivating EGFR signaling
    Authors: Wang, YY;Vadhan, A;Wu, CH;Hsu, CY;Chen, YC;Chen, YK;Chen, PY;Nguyen, HDH;Chang, YC;Yuan, SF;
    Oncology reports  2023-10-01
  18. Identification of a miRNA multi-targeting therapeutic strategy in glioblastoma
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    Cell death & disease  2023-09-25
  19. PLAC8-Mediated Activation of NOX4 Signalling Restores Angiogenic Function of Endothelial Colony-Forming Cells in Experimental Hypoxia
    Authors: Pun, SH;O'Neill, KM;Edgar, KS;Gill, EK;Moez, A;Naderi-Meshkin, H;Malla, SB;Hookham, MB;Alsaggaf, M;Madishetti, VV;Botezatu, B;King, W;Brunssen, C;Morawietz, H;Dunne, PD;Brazil, DP;Medina, RJ;Watson, CJ;Grieve, DJ;
    Cells  2023-09-06
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    Nature immunology  2023-09-04
  21. PTPN2 copper-sensing rapidly relays copper level fluctuations into EGFR/CREB activation and associated CTR1 transcriptional repression
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    bioRxiv : the preprint server for biology  2023-08-29
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    Authors: Wang, Y;Yang, C;Wang, Z;Wang, Y;Yan, Q;Feng, Y;Liu, Y;Huang, J;Zhou, J;
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    Cancers  2023-08-05
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    Nature communications  2023-08-04
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  26. Long-chain polyphosphates inhibit type I interferon signaling and augment LPS-induced cytokine secretion in human leukocytes
    Authors: Pirttiniemi, A;Adeshara, K;Happonen, N;Einarsdottir, E;Katayama, S;Salmenkari, H;H�rkk�, S;Kere, J;Groop, PH;Lehto, M;
    Journal of leukocyte biology  2023-05-24
  27. The role of KNTC1 in the regulation of proliferation, migration and tumorigenesis in colorectal cancer
    Authors: Wang, J;Ding, Z;Shu, W;Zhuge, Y;
    Cellular signalling  2023-05-23
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    Authors: Loda, A;Calza, S;Giacomini, A;Ravelli, C;Krishna Chandran, AM;Tobia, C;Tabellini, G;Parolini, S;Semeraro, F;Ronca, R;Rezzola, S;
    Cancer cell international  2023-05-11
  29. Targeting FGFRs Using PD173074 as a Novel Therapeutic Strategy in Cholangiocarcinoma
    Authors: Balasubramanian, B;Yacqub-Usman, K;Venkatraman, S;Myint, KZ;Juengsamarn, J;Sarkhampee, P;Lertsawatvicha, N;Sripa, J;Kuakpaetoon, T;Suriyonplengsaeng, C;Wongprasert, K;Grabowska, AM;Bates, DO;Janvilisri, T;Tohtong, R;
    Cancers  2023-04-28
  30. Galectin-3 promotes secretion of proteases that decrease epithelium integrity in human colon cancer cells
    Authors: S Li, DM Pritchard, LG Yu
    Cell Death & Disease, 2023-04-13;14(4):268.  2023-04-13
  31. Defining regorafenib as a senomorphic drug: therapeutic potential in the age-related lung disease emphysema
    Authors: JJ Park, K Oh, GW Lee, G Bang, JH Park, HB Kim, JY Kim, EY Shin, EG Kim
    Experimental & Molecular Medicine, 2023-04-03;0(0):.  2023-04-03
  32. IGFBP5 is an ROR1 ligand promoting glioblastoma invasion via ROR1/HER2-CREB signaling axis
    Authors: W Lin, R Niu, SM Park, Y Zou, SS Kim, X Xia, S Xing, Q Yang, X Sun, Z Yuan, S Zhou, D Zhang, HJ Kwon, S Park, C Il Kim, H Koo, Y Liu, H Wu, M Zheng, H Yoo, B Shi, JB Park, J Yin
    Nature Communications, 2023-03-22;14(1):1578.  2023-03-22
  33. Venetoclax-Resistant T-ALL Cells Display Distinct Cancer Stem Cell Signatures and Enrichment of Cytokine Signaling
    Authors: K Shah, L Al Ashiri, A Nasimian, M Ahmed, JU Kazi
    International Journal of Molecular Sciences, 2023-03-05;24(5):.  2023-03-05
  34. Stellettin B Induces Cell Death in Bladder Cancer Via Activating the Autophagy/DAPK2/Apoptosis Signaling Cascade
    Authors: CH Chang, BJ Lin, CH Chen, NL Nguyen, TH Hsieh, JH Su, MC Chen
    Marine Drugs, 2023-01-21;21(2):.  2023-01-21
  35. EGFR and Lyn inhibition augments regorafenib induced cell death in sorafenib resistant 3D tumor spheroid model
    Authors: E Sar?yar, O Karpat, S Sezan, SM Baylan, A K?pçak, K Guven, E Erdal, ZF Karagonlar
    Cellular Signalling, 2023-01-21;0(0):110608.  2023-01-21
  36. HIF-1alpha accumulation in response to transient hypoglycemia may worsen diabetic eye disease
    Authors: C Guo, M Deshpande, Y Niu, I Kachwala, M Flores-Bel, H Megarity, T Nuse, S Babapoor-F, M Ramada, J Sanchez, N Inamdar, TV Johnson, MV Canto-Sole, S Montaner, A Sodhi
    Cell Reports, 2023-01-10;42(1):111976.  2023-01-10
  37. IL-1RA promotes oral squamous cell carcinoma malignancy through mitochondrial metabolism-mediated EGFR/JNK/SOX2 pathway
    Authors: Yuan, S;Wang, Y;Chan, L;Hung, A;Nguyen, H;Chen, Y;Hu, S;Lo, S;Wang, Y;
    Research Square  2023-01-01
  38. CCL4 Regulates Eosinophil Activation in Eosinophilic Airway Inflammation
    Authors: HH Chu, Y Kobayashi, DV Bui, Y Yun, LM Nguyen, A Mitani, K Suzuki, M Asako, A Kanda, H Iwai
    International Journal of Molecular Sciences, 2022-12-18;23(24):.  2022-12-18
  39. Oxidized Proteins Differentially Affect Maturation and Activation of Human Monocyte-Derived Cells
    Authors: R Clemen, K Arlt, L Miebach, T von Woedtk, S Bekeschus
    Cells, 2022-11-18;11(22):.  2022-11-18
  40. BCG vaccinations drive epigenetic changes to the human T cell receptor: Restored expression in type 1 diabetes
    Authors: H Takahashi, WM Kühtreiber, RC Keefe, AH Lee, A Aristarkho, HF Dias, N Ng, KJ Nelson, S Bien, D Scheffey, DL Faustman
    Science Advances, 2022-11-16;8(46):eabq7240.  2022-11-16
  41. 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
  42. Extracellular Vesicles Isolated from Malignant Mesothelioma Cancer-Associated Fibroblasts Induce Pro-Oncogenic Changes in Healthy Mesothelial Cells
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    International Journal of Molecular Sciences, 2022-10-18;23(20):.  2022-10-18
  43. Loss of NECTIN1 triggers melanoma dissemination upon local IGF1 depletion
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  49. PTEN inhibits AMPK to control collective migration
    Authors: F Peglion, L Capuana, I Perfettini, L Boucontet, B Braithwait, E Colucci-Gu, E Quissac, K Forsberg-N, F Llense, S Etienne-Ma
    Nature Communications, 2022-08-11;13(1):4528.  2022-08-11
  50. p38 MAPK-dependent phosphorylation of transcription factor SOX2 promotes an adaptive response to BRAF inhibitors in melanoma cells
    Authors: S Pietrobono, R De Paolo, D Mangiameli, A Marranci, I Battisti, C Franchin, G Arrigoni, D Melisi, L Poliseno, B Stecca
    The Journal of Biological Chemistry, 2022-08-07;298(9):102353.  2022-08-07
  51. Angiotensin II Exaggerates SARS-CoV-2 Specific T-Cell Response in Convalescent Individuals following COVID-19
    Authors: M Almutlaq, FA Mansour, J Alghamdi, Y Alhendi, AA Alamro, AA Alghamdi, HS Alamri, F Alroqi, T Barhoumi
    International Journal of Molecular Sciences, 2022-08-04;23(15):.  2022-08-04
  52. Exosomes from EGFR-Mutated Adenocarcinoma Induce a Hybrid EMT and MMP9-Dependant Tumor Invasion
    Authors: A Jouida, M O'Callagha, C Mc Carthy, A Fabre, P Nadarajan, MP Keane
    Cancers, 2022-08-03;14(15):.  2022-08-03
  53. Tsp2 Facilitates Tumor-associated Fibroblasts Formation and Promotes Tumor Progression in Retroperitoneal Liposarcoma
    Authors: C Xu, L Yan, X Guan, Z Wang, J Wu, A Lv, D Liu, F Liu, B Dong, M Zhao, L Jia, X Tian, C Hao
    International journal of biological sciences, 2022-08-01;18(13):5038-5055.  2022-08-01
  54. The human amniotic epithelium confers a bias to differentiate toward the neuroectoderm lineage in human embryonic stem cells
    Authors: D Ávila-Gonz, W Portillo, CP Barragán-Á, G Hernandez-, E Flores-Gar, A Molina-Her, NE Díaz-Martí, NF Díaz
    Elife, 2022-07-11;11(0):.  2022-07-11
  55. Kayadiol exerted anticancer effects through p53-mediated ferroptosis in NKTCL cells
    Authors: C He, C Wang, H Liu, B Shan
    BMC Cancer, 2022-07-02;22(1):724.  2022-07-02
  56. Macrophage-derived CCL23 upregulates expression of T-cell exhaustion markers in ovarian cancer
    Authors: K Kamat, V Krishnan, O Dorigo
    British Journal of Cancer, 2022-06-24;0(0):.  2022-06-24
  57. Peroxiredoxin IV plays a critical role in cancer cell growth and radioresistance through the activation of the Akt/GSK3 signaling 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. 

View all Proteome Profiler Antibody Array FAQs

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Proteome Profiler Human Phospho-Kinase Array Kit
By Anonymous on 12/05/2024

Proteome Profiler Human Phospho-Kinase Array Kit
By Anonymous on 12/05/2024

Proteome Profiler Human Phospho-Kinase Array Kit
By Anonymous on 04/24/2023

Proteome Profiler Human Phospho-Kinase Array Kit
By Anonymous on 08/02/2022

Used human lymphoma samples! Worked fine!


Proteome Profiler Human Phospho-Kinase Array Kit
By Anonymous on 08/02/2022

Used this kit with mouse lymphoma cell lines and it worked!


Proteome Profiler Human Phospho-Kinase Array Kit
By Anonymous on 09/16/2021

Proteome Profiler Human Phospho-Kinase Array Kit
By Anonymous on 07/15/2021

Proteome Profiler Human Phospho-Kinase Array Kit
By Anonymous on 03/21/2021

Proteome Profiler Human Phospho-Kinase Array Kit
By Anonymous on 01/27/2021

Proteome Profiler Human Phospho-Kinase Array Kit
By APK Inmunometabolismo on 11/06/2020

Proteome Profiler Human Phospho-Kinase Array Kit
By Anonymous on 03/18/2020

Some 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


Proteome Profiler Human Phospho-Kinase Array Kit
By Anonymous on 01/21/2020