Kit Summary

A membrane-based antibody array for the parallel determination of the relative levels of mouse receptor tyrosine kinase phosphorylation. Validated for analyte detection in cell lysates.

Key Benefits

  • Detects phosphorylation of 39 mouse receptors simultaneously
  • Requires no specialized equipment

 

Principle of the Assay

The Proteome Profiler Mouse Phospho-RTK 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). Tyrosine phosphorylation of the captured proteins is detected with an HRP-conjugated pan phospho-tyrosine antibody (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 receptors 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
  • 4 Array Membranes
  • 4-Well Multi-dish
  • Array Buffers
  • Lysis Buffer
  • Wash Buffer
  • Anti-Phospho-Tyrosine-HRP Detection Antibody
  • Chemiluminescent Detection Reagents
  • Transparency Overlay Template
  • Detailed 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 RTKs in a single sample
EGF R PDGF R alpha VEGF R3
ErbB2 PDGF R beta MuSK
ErbB3 SCF R EphA1
ErbB4 Flt-3 EphA2
FGF R2 (IIIc) M-CSF R EphA3
FGF R3 c-Ret EphA6
FGF R4 Tie-1 EphA7
Insulin R Tie-2 EphA8
IGF-I R TrkA EphB1
Axl TrkB EphB2
DtK TrkC EphB4
Mer VEGF R1 EphB6
HGF R VEGF R2  
MSP R    

 

Assays for analytes represented in the Mouse Phospho-Receptor Tyrosine Kinase Array Kit

  DuoSet® ELISA Development Reagents DuoSet® IC ELISA Development Reagents (Total) DuoSet® IC ELISA Development Reagents (Phospho) Quantikine® ELISA Kits Cell-based ELISA Kits
EGF R          
ErbB2          
ErbB3          
ErbB4          
FGF R2 (IIIc)          
FGF R3          
FGF R4          
Insulin R          
IGF-I R          
Axl DY854        
DtK DY759        
Mer DY591        
HGF R DY527        
MSP R          
PDGF R alpha          
PDGF R beta          
SCF R          
Flt-3          
M-CSF R          
c-Ret          
Tie-1          
Tie-2     DYC2816    
TrkA          
TrkB          
TrkC          
VEGF R1 DY471     MVR100  
VEGF R2 DY1558B     MVR200B  
VEGF R3 DY743        
MuSK          
EphA1          
EphA2          
EphA3          
EphA6          
EphA7          
EphA8          
EphB1          
EphB2          
EphB4          
EphB6          

Troubleshooting Guide

Data Examples

Figure 1.
View Larger Image

Figure 1: The Mouse Phospho-RTK Array detects multiple tyrosine phosphorylated receptors in untreated and ligand-treated cell lysates. The amount of lysate incubated with each array is indicated in the figure. Data shown are from 2 minute (A, B, and C) or 5 minute (D) exposure to X-ray film.

A. M1 mouse myeloid leukemia cells were either untreated or treated with 500 ng/mL recombinant mouse Flt-3 Ligand (Catalog # 427-FL) for 5 minutes.

B. NMuMG mouse mammary gland epithelial cells were either untreated or treated with 200 ng/mL recombinant mouse EGF (Catalog # 2028-EG) for 5 minutes.

C. Hepa 1-6 mouse hepatoma cells were either untreated or treated with 1 µg/mL recombinant human insulin (Sigma, Catalog # I9278).

D. HEK293 human embryonic kidney cells transfected with mouse EphA1 were either untreated or treated with 3 µg/mL mouse Ephrin-A1 (Catalog # 602-A1) and 0.3 µg/mL goat anti-human IgG Fc (Catalog # G-102-C) for 20 minutes.

Preparation and Storage
  • Stability & Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: RTK Assay Kits
Receptor Tyrosine Kinases (RTKs) are widely expressed transmembrane proteins that act as receptors for growth factors, neurotrophic factors, and other extracellular signaling molecules. Upon ligand binding, they undergo tyrosine phosphorylation at specific residues in the cytoplasmic tail. This leads to the binding of protein substrates and/or the establishment docking sites for adaptor proteins involved in RTK-mediated signal transduction. RTKs have critical functions in several developmental processes including regulating cell survival, proliferation, and motility. When unregulated, they play prominent roles in cancer formation. R&D Systems offers several different multiplex assay formats that allow the user to assess the activity of many RTKs in a single sample. Available RTK multiplex kits include bead-based multiplex kits for the Luminex platform, membrane-based antibody arrays, and microplate-based antibody arrays.
  • Alternate Names:
    RTK Assay Kits
Related Research Areas
Citations:

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.

15 Citations: Showing 1 - 10
Filter your results:

Species
Sample Type
  1. Enzymatic cleavage of myoferlin releases a dual C2-domain module linked to ERK signalling
    Authors: AK Piper, SE Ross, GM Redpath, FA Lemckert, N Woolger, A Bournazos, PA Greer, RB Sutton, ST Cooper
    Cell. Signal, 2017;33(0):30-40. 2017
  2. HBEGF promotes gliomagenesis in the context of Ink4a/Arf and Pten loss
    Authors: CH Shin, JP Robinson, JA Sonnen, AE Welker, DX Yu, MW VanBrockli, SL Holmen
    Oncogene, 2017;0(0):. 2017
  3. Gemcitabine-induced TIMP1 attenuates therapy response and promotes tumor growth and liver metastasis in pancreatic cancer
    Authors: Z D'Costa, K Jones, A Azad, R van Stipho, SY Lim, AL Gomes, P Kinchesh, SC Smart, WG McKenna, FM Buffa, OJ Sansom, RJ Muschel, E O'Neill, E Fokas
    Cancer Res., 2017;0(0):. 2017
  4. OCD-like behavior is caused by dysfunction of thalamo-amygdala circuits and upregulated TrkB/ERK-MAPK signaling as a result of SPRED2 deficiency
    Authors: M Ullrich, M Weber, AM Post, S Popp, J Grein, M Zechner, H Guerrero G, A Kreis, AG Schmitt, N Üçeyler, KP Lesch, K Schuh
    Mol. Psychiatry, 2017;0(0):. 2017
  5. Insulin-mediated signaling facilitates resistance to PDGFR inhibition in proneural hPDGFB-driven gliomas
    Authors: DA Almiron Bo, C Ran, MC Havrda, H Liu, Y Hitoshi, Z Zhang, C Cheng, M Ung, MA Israel
    Mol. Cancer Ther, 2017;0(0):. 2017
  6. Epidermal growth factor receptor inhibition reduces angiogenesis via hypoxia-inducible factor-1alpha and Notch1 in head neck squamous cell carcinoma.
    Authors: Wang W, Zhao Z, Ma S, Yu G, Liu B, Zhang L, Zhang W, Kulkarni A, Sun Z, Zhao Y
    PLoS ONE, 2015;10(2):e0119723. 2015
  7. Lack of Matrilin-2 favors liver tumor development via Erk1/2 and GSK-3beta pathways in vivo.
    Authors: Fullar A, Baghy K, Deak F, Peterfia B, Zsak Y, Tatrai P, Schaff Z, Dudas J, Kiss I, Kovalszky I
    PLoS ONE, 2014;9(4):e93469. 2014
  8. ZEB1 sensitizes lung adenocarcinoma to metastasis suppression by PI3K antagonism.
    Authors: Yang Y, Ahn Y, Chen Y, Tan X, Guo L, Gibbons D, Ungewiss C, Peng D, Liu X, Lin S, Thilaganathan N, Wistuba I, Rodriguez-Canales J, McLendon G, Creighton C, Kurie J
    J Clin Invest, 2014;124(6):2696-708. 2014
  9. HPV16-associated tumors control myeloid cell homeostasis in lymphoid organs, generating a suppressor environment for T cells.
    Authors: Stone S, Rossetti R, Bolpetti A, Boccardo E, Souza P, Lepique A
    J Leukoc Biol, 2014;96(4):619-31. 2014
  10. Decorin deficiency promotes hepatic carcinogenesis.
    Authors: Horvath Z, Kovalszky I, Fullar A, Kiss K, Schaff Z, Iozzo R, Baghy K
    Matrix Biol, 2014;35(0):194-205. 2014
  11. Erlotinib prolongs survival in pancreatic cancer by blocking gemcitabine-induced MAPK signals.
    Authors: Miyabayashi K, Ijichi H, Mohri D, Tada M, Yamamoto K, Asaoka Y, Ikenoue T, Tateishi K, Nakai Y, Isayama H, Morishita Y, Omata M, Moses H, Koike K
    Cancer Res, 2013;73(7):2221-34. 2013
  12. Deletion of the epidermal growth factor receptor in renal proximal tubule epithelial cells delays recovery from acute kidney injury.
    Authors: Chen J, Chen JK, Harris RC
    Kidney Int., 2012;82(1):45-52. 2012
  13. Lentiviral vector induced insertional haploinsufficiency of Ebf1 causes murine leukemia.
    Authors: Heckl D, Schwarzer A, Haemmerle R, Steinemann D, Rudolph C, Skawran B, Knoess S, Krause J, Li Z, Schlegelberger B, Baum C, Modlich U
    Mol. Ther., 2012;20(6):1187-95. 2012
  14. Estrogenic compounds are not always cardioprotective and can be lethal in males with genetic heart disease.
    Authors: Haines, Christop, Harvey, Pamela A, Luczak, Elizabet, Barthel, Kristen, Konhilas, John P, Watson, Peter A, Stauffer, Brian L, Leinwand, Leslie A
    Endocrinology, 2012;153(9):4470-9. 2012
  15. Specific glycosaminoglycans modulate neural specification of mouse embryonic stem cells.
    Authors: Pickford CE, Holley RJ, Rushton G, Stavridis MP, Ward CM, Merry CL
    Stem Cells, 2011;29(4):629-40. 2011
Expand to show all 15 Citations
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Average Rating: 5 (Based on 3 reviews)

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Images Ratings Applications Species Reviewed By Date Details
 Proteome Profiler Mouse Phospho-RTK Array Kit ARY014
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Excellent
  Anonymous 07/05/2017
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 Proteome Profiler Mouse Phospho-RTK Array Kit ARY014
: Proteome Profiler Mouse Phospho-RTK Array Kit [ARY014]

Other Experimental Details

Other Experimental DetailsLysates from mouse embryonic stem cells applied to membranes using instructions provided. Western blots were clean and worked great!
 Proteome Profiler Mouse Phospho-RTK Array Kit ARY014
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Excellent
  Anonymous 05/16/2017
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 Proteome Profiler Mouse Phospho-RTK Array Kit ARY014
: Proteome Profiler Mouse Phospho-RTK Array Kit [ARY014]

Other Experimental Details

Other Experimental DetailsWorked really well on mouse tumour lysates.
 Proteome Profiler Mouse Phospho-RTK Array Kit ARY014
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Excellent
  Anonymous 06/29/2016
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 Proteome Profiler Mouse Phospho-RTK Array Kit ARY014
: Proteome Profiler Mouse Phospho-RTK Array Kit [ARY014]

Other Experimental Details

Other Experimental DetailsMammary tumors extracts from MMTV-Neu mice with different p53 genotypes (p53wt vs mutp53)

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