Proteome Profiler Human Phospho-Immunoreceptor Array Kit ARY004B: R&D Systems
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  

 

Assays for analytes represented in the Human Phospho-Kinase Array Kit

Analyte DuoSet® IC ELISA Development Systems (Total) DuoSet® IC ELISA
Development Systems (Phospho)
Surveyor™ IC or Quantikine®
ELISA Kits
Cell-based
ELISA Kits
Akt (S473)   DYC887B SUV887B  
Akt (T308)        
AMPK alpha1 (T174) DYC3197 DYC3528    
AMPK alpha2 (T172)        
beta-Catenin DYC1329   DYC1329  
Chk2 (T68)   DYC1626    
c-Jun (S63)       KCB7499
CREB (S133)   DYC2510    
EGF R (Y1068) DYC1854 DYC3570 DEGFR0  
eNOS (S1177)        
ERK1/2 (T202/Y204, T185/Y187)   DYC1018B    
FAK (Y397) DYC4467      
Fgr (Y412)        
Fyn (Y420)        
GSK-3 alpha/beta (S21/S9) DYC2157 DYC2630    
Hck (Y411)        
HSP27 (S78/S82) DYC1580 DYC2314    
HSP60 DYC1800      
JNK pan (T183/Y185, T221/Y223) DYC1205 DYC1387    
Lck (Y394)        
Lyn (Y397)   DYC3936    
MSK1/2 (S376/S360)        
p27 (T198) DYC2256      
p38 alpha (T180/Y182) DYC8691B DYC869B    
p53 (S15) DYC1043 DYC1839    
p53 (S46) DYC1043 DYC1489    
p53 (S392) DYC1043 DYC2996    
p70 S6 Kinase (T421/S424) DYC8962 DYC8965    
PDGF R beta (Y751) DYC385 DYC3096    
PLC gamma-1 (Y783)        
PRAS40 (T246)   DYC6890    
PYK2 (Y402)        
RSK1/2/3 (S380/S386/S377)   DYC889B    
STAT2 (Y689)        
STAT3 (S727)        
STAT3 (Y705)   DYC4607B    
STAT5a (Y694)        
STAT5a/b (Y694/Y699)        
STAT5b (Y699)        
STAT6 (Y641)        
TOR (S2448)        
WNK-1(T60)   DYC4720    
Yes (Y426)   DYC3929    

 

Troubleshooting Guide

 

Data Examples
Figure 1A.
View Larger Image

Figure 1A: The Human Phospho-Immunoreceptor Array detects multiple tyrosine phosphorylated immunoreceptors in lysates prepared from cells activated by antibodymediated cross-linking of cell-surface immunoreceptors. THP-1 human acute monocytic leukemia cells were incubated with goat anti-Fcγ 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].

Figure 1B.
View Larger Image

Figure 1B: Jurkat human acute T cell leukemia cells were incubated with a mouse monoclonal anti-CD3ε 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.

Preparation and Storage
  • Stability & 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.
  • Alternate Names:
    Immunoreceptor Array Kit
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.

3 Citations: Showing 1 - 3
Filter your results:

Species
Sample Type
  1. Evidence for C1q-mediated crosslinking of CD33/LAIR-1 inhibitory immunoreceptors and biological control of CD33/LAIR-1 expression
    Authors: M Son, B Diamond, BT Volpe, CB Aranow, MC Mackay, F Santiago-S
    Sci Rep, 2017;7(1):270. 2017
  2. Fibroblast Growth Factor Receptor 3 (FGFR3) Associated with the CD20 Antigen Regulates the Rituximab-induced Proliferation Inhibition in B-cell Lymphoma Cells.
    Authors: Kotani N, Ishiura Y, Yamashita R, Ohnishi T, Honke K
    J Biol Chem, 2012;287(44):37109-18. 2012
  3. Surface molecule CD229 as a novel target for the diagnosis and treatment of multiple myeloma.
    Authors: Atanackovic D, Panse J, Hildebrandt Y, Jadczak A, Kobold S, Cao Y, Templin J, Meyer S, Reinhard H, Bartels K, Lajmi N, Zander AR, Marx AH, Luetkens T, Bokemeyer C, Kroger N
    Haematologica, 2011;0(0):. 2011
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