Proteome Profiler Mouse Phospho-RTK Array Kit
Proteome Profiler Mouse Phospho-RTK Array 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.
- 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.
- 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|
|FGF R2 (IIIc)||M-CSF R||EphA3|
|HGF R||VEGF R2|
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|
|FGF R2 (IIIc)|
|PDGF R alpha|
|PDGF R beta|
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
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.
Citations for Proteome Profiler Mouse Phospho-RTK 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.
Citations: Showing 1 - 10
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EphB2-dependent signaling promotes neuronal excitotoxicity and inflammation in the acute phase of ischemic stroke
Authors: AS Ernst, LI Böhler, AM Hagenston, A Hoffmann, S Heiland, C Sticht, M Bendszus, M Hecker, H Bading, HH Marti, T Korff, R Kunze
Acta Neuropathol Commun, 2019;7(1):15. 2019
KSHV-induced ligand mediated activation of PDGF receptor-alpha drives Kaposi's sarcomagenesis
Authors: LE Cavallin, Q Ma, J Naipauer, S Gupta, M Kurian, P Locatelli, P Romanelli, M Nadji, PJ Goldschmid, EA Mesri
PLoS Pathog., 2018;14(7):e1007175. 2018
Glycosylation controls cooperative PECAM-VEGFR2-?3 integrin functions at the endothelial surface for tumor angiogenesis
Authors: R Imamaki, K Ogawa, Y Kizuka, Y Komi, S Kojima, N Kotani, K Honke, T Honda, N Taniguchi, S Kitazume
Oncogene, 2018;0(0):. 2018
Heat shock factor 1 confers resistance to lapatinib in ERBB2-positive breast cancer cells
Authors: A Yallowitz, A Ghaleb, L Garcia, EM Alexandrov, N Marchenko
Cell Death Dis, 2018;9(6):621. 2018
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
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
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
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
Betacellulin regulates the proliferation and differentiation of retinal progenitor cells in vitro
Authors: D Zhang, B Shen, Y Zhang, N Ni, Y Wang, F Xianqun, H Sun, P Gu
J. Cell. Mol. Med., 2017;0(0):. 2017
Effective combinatorial immunotherapy for castration-resistant prostate cancer
Authors: X Lu, JW Horner, E Paul, X Shang, P Troncoso, P Deng, S Jiang, Q Chang, DJ Spring, P Sharma, JA Zebala, DY Maeda, YA Wang, RA DePinho
Nature, 2017;543(7647):728-732. 2017
Axonal outgrowth, neuropeptides expression and receptors tyrosine kinase phosphorylation in 3D organotypic cultures of adult dorsal root ganglia
Authors: E Neto, CJ Alves, L Leitão, DM Sousa, IS Alencastre, F Conceição, M Lamghari
PLoS ONE, 2017;12(7):e0181612. 2017
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
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
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
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
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
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
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
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
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
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
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
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Reviews for Proteome Profiler Mouse Phospho-RTK Array Kit
Average Rating: 5 (Based on 5 Reviews)
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Lysates from mouse embryonic stem cells applied to membranes using instructions provided. Western blots were clean and worked great!
Worked really well on mouse tumour lysates.