R&D Systems Human Phospho-Immunoreceptor Array (Catalog # ARY004) is a rapid, sensitive, and economical tool used to simultaneously detect the relative phosphorylation levels of 59 ITAM/ITIM-associated immunoreceptors, avoiding numerous immunoprecipitations and/or Western blots.
Proteome Profiler Human Phospho-Immunoreceptor Array Kit
R&D Systems | Catalog # ARY004B
Key Product Details
Species
Product Summary for Proteome Profiler Human Phospho-Immunoreceptor Array Kit
Kit Summary
A membrane-based antibody array for the parallel determination of the relative phosphorylation of human immunoreceptors. Validated for analyte detection in cell lysates.
Key Benefits
- Detects phosphorylation of 59 human ITAM/ITIM-associated immunoreceptors simultaneously
- Requires no specialized equipment
Principle of the Assay
The Proteome Profiler Human Phospho-Immunoreceptor 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 immunoreceptors 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
This kit contains the following azide-free, fluorochrome-conjugated antibodies to verify human stem cell pluripotency:
- 4 Array Membranes
- 4-Well Multi-dish
- Array Buffers
- Lysis Buffer
- Wash Buffer
- HRP-conjugated Anti-phospho-tyrosine 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.
| Simultaneously detect the relative phosphorylation levels of these immunoreceptors in a single sample. | ||
|---|---|---|
| 2B4/SLAMF4 | FcRH1/IRTA5 | NKp46/NCR1 |
| BLAME/SLAMF8 | FcRH2/IRTA4 | NKp80/KLRF1 |
| BTLA | FcRH4/IRTA1 | NTB-A/SLAMF6 |
| CD3 epsilon | FcRH5/IRTA2 | PD-1 |
| CD5 | ILT2/CD85j | PECAM/CD31) |
| CD6 | ILT3/CD85k | SHIP-1 |
| CD28 | ILT4/CD85d | SHP-1 |
| CD84/SLAMF5 | ILT5/CD85a | SHP-2 |
| CD229/SLAMF3 | ILT6/CD85e | Siglec-2/CD22 |
| CEACAM-1 | Integrin beta 3/CD61 | Siglec-3/CD33 |
| CLEC-1 | KIR2DL4 | Siglec-5 |
| CLEC-2 | LAIR-1 | Siglec-7 |
| CRACC/SLAMF7 | LAIR-2 | Siglec-9 |
| CTLA-4/CD152 | LMIR1/CD300A | Siglec-10 |
| DCIR/CLEC4A | LMIR2/CD300C | SIRP-beta 1 |
| Dectin-1/CLEC7A | LMIR3/CD300LF | SLAM/CD150 |
| DNAM-1 | LMIR6/CD300LE | TREM-1 |
| Fc epsilon RII/CD23 | MDL-1/CLEC5A | TREM-2 |
| Fc gamma RIIA | NKp30/NCR3 | TREML1/TLT-1 |
| Fc gamma RIIIA/B | NKp44/NCR2 | |
Scientific Data Images for Proteome Profiler Human Phospho-Immunoreceptor Array Kit
The Human Phospho-Immunoreceptor Array detects multiple tyrosine phosphorylated immunoreceptors in lysates prepared from cells activated by antibody mediated cross-linking of cell-surface immunoreceptors.
Jurkat human acute T cell leukemia cells were incubated with a mouse monoclonal anti-CD3 epsilon antibody (R&D Systems, Catalog # MAB100) or mouse IgG1 isotype control (R&D Systems, Catalog # MAB002) followed by incubation with a goat anti-mouse antibody (R&D Systems, Catalog # AF007) for 5 minutes. 100 µg of lysate was run on each array.
The Human Phospho-Immunoreceptor Array detects multiple tyrosine phosphorylated immunoreceptors in lysates prepared from cells activated by antibody mediated cross-linking of cell-surface immunoreceptors.
THP-1 human acute monocytic leukemia cells were incubated with goat anti-Fc gamma RIIA antibody (R&D Systems, Catalog # AF1875) or normal goat IgG (R&D Systems, Catalog # AB-108-C) followed by incubation with a donkey anti-goat IgG antibody (R&D Systems, Catalog # AF109) for 5 minutes [Maresco, D.L. et al. (1999) J. Immunol. 162:6458].
Formulation, Preparation, and Storage
Shipping
Storage
Background: Immunoreceptor Array Kit
Additional Immunoreceptor Array Kit Products
Product Documents for Proteome Profiler Human Phospho-Immunoreceptor Array Kit
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Proteome Profiler Human Phospho-Immunoreceptor Array Kit
For research use only
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Citations for Proteome Profiler Human Phospho-Immunoreceptor Array Kit
Customer Reviews for Proteome Profiler Human Phospho-Immunoreceptor Array Kit (1)
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Verified Customer | Posted 09/23/2021very good product, easy protocol, good results. it can be kept in 4-degree fridge.
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FAQs for Proteome Profiler Human Phospho-Immunoreceptor Array Kit
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Q: The insert states to lyse 1 x 107 cells in 1 mL Lysis Buffer 17. What is the approximate protein concentration of the resulting lysate?
A: Lysing following recommendations in the product insert produces a lysate with a protein concentration of about 1.5-2.0 mg/mL. Quantitation of sample protein concentration using a total protein assay is recommended.