Proteome Profiler Human Phospho-Immunoreceptor Array Kit

R&D Systems | Catalog # ARY004B

Contains 4 membranes - each spotted in duplicate with 59 different immunoreceptor antibodies
R&D Systems
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Key Product Details

Species

Human

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.

Troubleshooting Guide

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  
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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.

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.

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

The product is shipped with polar packs. 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.

Background: Immunoreceptor Array Kit

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.

Additional Immunoreceptor Array Kit Products

Product Documents for Proteome Profiler Human Phospho-Immunoreceptor Array Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Proteome Profiler Human Phospho-Immunoreceptor Array Kit

For research use only

Citations for Proteome Profiler Human Phospho-Immunoreceptor Array Kit

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  • Proteome Profiler Human Phospho-Immunoreceptor Array Kit
    Name: Anonymous
    Verified Customer | Posted 09/23/2021
    very good product, easy protocol, good results. it can be kept in 4-degree fridge.
    Proteome Profiler Human Phospho-Immunoreceptor Array Kit ARY004B

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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.
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