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.
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.
- 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 |
Specifications
Product Datasheets
Scientific Data
Detection of Multiple Tyrosine Phosphorylated Receptors in Cell Lysates by the Mouse Phospho-RTK Array. The amount of lysate incubated with each array is indicated in the figure. Data shown are from 2 minute (Panels A, B, and C) or 5 minute (Panel 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.
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.
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BioID-based intact cell interactome of the Kv1.3 potassium channel identifies a Kv1.3-STAT3-p53 cellular signaling pathway
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KSHV-induced ligand mediated activation of PDGF receptor-alpha drives Kaposi's sarcomagenesis
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FAQs
-
What are the phosphorylation site(s) of the 49 different tyrosine residues on the RTKs?
This kit uses a pan anti-phospho-tyrosine antibody as the detection antibody, which means it is capable of detecting phosphorylation at any available tyrosine. It is not designed to be specific for one phosphorylation site on each molecule. For instance, PDGF R alpha has multiple sites of tyrosine phosphorylation (see the “PTM/Processing” section of http://www.uniprot.org/uniprot/P16234 for a list of the currently identified sites).
-
Can the Mouse Phospho-RTK Array be used for measuring the relative levels of total RTKs present in a sample?
No. Although the array kit uses capture antibodies that recognize both phosphorylated and unphosphorylated RTKs, the detection antibody is a pan anti-phospho-tyrosine antibody which only detects phosphorylated tyrosines on activated RTKs.
-
To confirm positive signal for an RTK by IP-Western blot, are the capture antibodies from the Mouse Phospho-RTK Array Kit offered separately?
The identities of the capture antibodies on the Mouse Phospho-RTK Array are considered proprietary. However, to help investigators confirm their array results, Bio-Techne offers antibodies for each RTK detectable in the array kit. These antibodies are validated for Western blot and can be found in our catalog. A pan anti-phospho-tyrosine-HRP detection antibody is also available (Catalog # HAM1676).
Reviews for Proteome Profiler Mouse Phospho-RTK Array Kit
Average Rating: 4.5 (Based on 8 Reviews)
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Use of RTK array to assay FACS sorted cells from murine lungs.
Sample from FACS sorted cells, so limited amount of protein but still able to see some phospho RTK. Would definitely use again.
Array works and produces publishable results. Not happy with the pho-PDGFR antibody, sinse it appear to cross react with something in mouse tumor lysate
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.