Proteome Profiler Mouse XL Cytokine Array ARY028: R&D Systems
Kit Summary

A membrane-based antibody array for the parallel determination of the relative levels of selected human cytokines and chemokines. Validated for analyte detection in cell culture supernates, cell lysates, tissue lysates, serum, and urine.

Key Benefits

  • Detects 111 mouse cytokines simultaneously
  • Requires no specialized equipment
  • Compatible with LI-COR and chemiluminescence detection


LI-COR Data Examples for
Mouse XL Cytokine Array Kit
 

 

Principle of the Assay

The Proteome Profiler Mouse XL Cytokine 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). Captured proteins are detected with biotinylated detection antibodies (Step 2) and then visualized using chemiluminescent detection reagents (Step 3). The signal produced is proportional to the amount of analyte bound.

Why Use an Antibody Array to Detect Multiple Cytokines?

Determining the expression of multiple cytokines 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 cytokines 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:

  • Rectangular 4-Well Multi-dish
  • 4 Mouse XL Cytokine Array nitrocellulose membranes spotted with 111 different antibodies to mouse cytokines
  • Array Buffer 4
  • Array Buffer 6
  • Chemi Reagent 1
  • Chemi Reagent 2
  • Detection Antibody Cocktail, Mouse XL Cytokine Array
  • Streptavidin-HRP
  • Transparency Overlay Template
  • Wash Buffer Concentrate (25X)

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 levels of these cytokines, chemokines, and growth factors in a single sample.
Adiponectin/Acrp30 DPPIV/CD26 IL-27
Amphiregulin EGF IL-28
Angiopoietin-1 Endoglin/CD105 IL-33
Angiopoietin-2 Endostatin LDL R
Angiopoietin-like 3 Fetuin A/AHSG Leptin
BAFF/BLyS/TNFSF13B FGF acidic LIF
C1q R1/CD93 FGF-21 Lipocalin-2/NGAL,
CCL2/JE/MCP-1 Flt-3 Ligand LIX
CCL3/CCL4 MIP-1 alpha/beta Gas6 M-CSF
CCL5/RANTES G-CSF MMP-2
CCL6/C10 GDF-15 MMP-3
CCL11/Eotaxin GM-CSF MMP-9
CCL12/MCP-5 HGF Myeloperoxidase
CCL17/TARC ICAM-1/CD54 Osteopontin (OPN)
CCL19/MIP-3 beta IFN-gamma Osteoprotegerin/TNFRSF11B
CCL20/MIP-3 alpha IGFBP-1 PD-ECGF/Thymidine phosphorylase
CCL21/6Ckine IGFBP-2 PDGF-BB
CCL22/MDC IGFBP-3 Pentraxin 2/SAP
CD14 IGFBP-5 Pentraxin 3/ TSG-14
CD40/TNFRSF5 IGFBP-6 Periostin/OSF-2
CD160 IL-1 alpha/IL1F1 Pref-1/DLK-1/FA1
Chemerin IL-1 beta/IL-1F2 Proliferin
Chitinase 3-like 1 IL-1ra/IL-1F3 Proprotein Convertase 9/PCSK9
Coagulation Factor III/Tissue Factor IL-2 RAGE
Complement Component C5/C5a IL-3 RBP4
Complement Factor D IL-4 Reg3G
C-Reactive Protein/CRP IL-5 Resistin
CX3CL1/Fractalkine IL-6 E-Selectin/CD62E
CXCL1/KC IL-7 P-Selectin/CD62P
CXCL2/MIP-2 IL-10 Serpin E1/PAI-1
CXCL9/MIG IL-11 Serpin F1/PEDF
CXCL10/IP-10 IL-12p40 Thrombopoietin
CXCL11/I-TAC IL-13 TIM-1/KIM-1/HAVCR
CXCL13/BLC/BCA-1 IL-15 TNF-alpha
CXCL16 IL-17A VCAM-1/CD106
Cystatin C  IL-22 VEGF
Dkk-1 IL-23 WISP-1/CCN4

Assays for Analytes represented in the Mouse XL Cytokine Array Kit

Analyte Qkit Duoset
Adiponectin/Acrp30 MRP300   DY1119  
Amphiregulin     DY989  
Angiopoietin-1        
Angiopoietin-2        
Angiopoietin-like 3 MANL30   DY136  
BAFF/BLyS/TNFSF13B MBLYS0   DY2106-05  
C1q R1/CD93 MCD930      
CCL2/JE/MCP-1 MJE00   DY479  
CCL3/CCL4 MIP-1 alpha/beta        
CCL5/RANTES MMR00   DY478  
CCL6/C10     DY487  
CCL11/Eotaxin MME00   DY420  
CCL12/MCP-5 MCC120   DY428  
CCL17/TARC MCC170   DY529  
CCL19/MIP-3 beta     DY440  
CCL20/MIP-3 alpha MCC200   DY760  
CCL21/6Ckine     DY457  
CCL22/MDC MCC220   DY439  
CD14 MC140   DY982  
CD40/TNFRSF5 MCCD40   DY097  
CD160 MCHM00   DY2325  
Chemerin        
Chitinase 3-like 1 MC3L10   DY2649  
Coagulation Factor III/Tissue Factor        
Complement Component C5/C5a        
Complement Factor D        
C-Reactive Protein/CRP MCRP00   DY1829  
CX3CL1/Fractalkine MCX310   DY472  
CXCL1/KC MKC00B   DY453  
CXCL2/MIP-2 MM200   DY452  
CXCL9/MIG MCX900   DY492  
CXCL10/IP-10 MCX100   DY466  
CXCL11/I-TAC     DY572  
CXCL13/BLC/BCA-1 MCX130      
CXCL16     DY503  
Cystatin C  MSCTC0   DY1238  
Dkk-1 MKK100   DY1765  
DPPIV/CD26     DY954  
EGF MEG00   DY2028  
Endoglin/CD105 MNDG00   DY1320  
Endostatin        
Fetuin A/AHSG MFTA00   DY1563  
FGF acidic     DY4686-05  
FGF-21 MF2100   DY3057  
Flt-3 Ligand MFK00   DY427  
Gas6 MGAS60   DY986  
G-CSF MCS00   DY414  
GDF-15 MGD150   DY6385  
GM-CSF MGM00   DY415  
HGF MHG00   DY2207  
ICAM-1/CD54 MIC100   DY796  
IFN-gamma MIF00   DY485  
IGFBP-1     DY1588-05  
IGFBP-2     DY797  
IGFBP-3 MGB300   DY775  
IGFBP-5     DY578  
IGFBP-6     DY776  
IL-1 alpha/IL1F1 MLA00   DY400  
IL-1 beta/IL-1F2 MLB00C   DY401  
IL-1ra/IL-1F3 MRA00   DY480  
IL-2 M2000   DY402  
IL-3 M3000   DY403  
IL-4 M4000B   DY404  
IL-5 M5000   DY405  
IL-6 M6000B   DY406  
IL-7 M7000   DY407  
IL-10 M1000B   DY417  
IL-11     DY418  
IL-12p40 MP400
non-allele specific
M1240
allele specific
DY2398
non-allele specific
DY499
allele specific
IL-13 M1300CB   DY413  
IL-15     DY447  
IL-17A M1700   DY421  
IL-22 M2200   DY582  
IL-23 M2300   DY1887  
IL-27     DY1789B  
IL-28        
IL-33 M3300   DY3626  
LDL R MLDLR0   DY2255  
Leptin MOB00   DY498  
LIF MLF00      
Lipocalin-2/NGAL,     DY1857  
LIX MX000   DY443  
M-CSF MMC00   DY416  
MMP-2 MMP200      
MMP-3 MMP300   DY548  
MMP-9 MMPT90 MMP900B DY6718 DY909
Myeloperoxidase     DY3667  
Osteopontin (OPN) MOST00   DY441  
Osteoprotegerin/TNFRSF11B MOP00   DY459  
PD-ECGF/Thymidine phosphorylase        
PDGF-BB MBB00      
Pentraxin 2/SAP MPTX20      
Pentraxin 3/ TSG-14 MPTX30   DY2166  
Periostin/OSF-2 MOSF20   DY2955  
Pref-1/DLK-1/FA1        
Proliferin        
Proprotein Convertase 9/PCSK9 MPC900   DY3985  
RAGE MRG00   DY1179  
RBP4 MRBP40   DY3476  
Reg3G        
Resistin MRSN00   DY1069  
E-Selectin/CD62E MES00   DY575  
P-Selectin/CD62P MPS00   DY737  
Serpin E1/PAI-1     DY3828-05  
Serpin F1/PEDF        
Thrombopoietin MTP00   DY488  
TIM-1/KIM-1/HAVCR MKM100   DY1817  
TNF-alpha MTA00B   DY410  
VCAM-1/CD106 MVC00   DY643  
VEGF MMV00   DY493  
WISP-1/CCN4 MWSP10      
 

 

Data Examples
Figure 1.
View Larger Image

Figure 1. Lysates from Balb/3T3 mouse embryonic fibroblast cells were untreated or treated with 100 ng/mL of recombinant mouse TNF-alpha (R&D Systems, Catalog # 410-MT) for 24 hours (200 µg lysate, 10 minute exposure).

Figure 2.
View Larger Image

Figure 2. Lysates from CMT-93 mouse rectal carcinoma cells were untreated or treated with 100 ng/mL LPS for 24 hours (200 µg lysate, 10 minute exposure).

Figure 3.
View Larger Image

Figure 3. Balb/3T3 mouse embryonic fibroblast cells were untreated or treated with 100 ng/mL of recombinant mouse TNF-alpha (R&D Systems, Catalog # 410-MT) for 24 hours (500 µL of cell culture supernate, 5 minute exposure).

Preparation and Storage
  • Stability & Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Assay Procedure

Refer to the product datasheet for complete product details.

Briefly, human stem cell pluripotency can be verified in live cells prior to colony selection or experimentation using this 30 minute procedure:

  • Pluripotent stem cell marker antibodies are added directly to the cells of interest
  • After 30 minutes, the cells are washed and analyzed for the expression of pluripotency markers
  • Positive colonies can be selected
  • Prepare membrane and incubate with prepared sample
  • Incubate the membrane with Detection Antibody Cocktail
  • Incubate the membrane array with Streptavidin-HRP
  • Develop the membrane array with Chemi Reagents 1 and 2
  • Expose the membrane array to autoradiography film
  • for expansion in culture
 

 

Kit Contents
  • Rectangular 4-Well Multi-dish
  • 4 Mouse XL Cytokine Array nitrocellulose membranes spotted with 111 different antibodies to mouse cytokines
  • Array Buffer 4
  • Array Buffer 6
  • Chemi Reagent 1
  • Chemi Reagent 2
  • Detection Antibody Cocktail, Mouse XL Cytokine Array
  • Streptavidin-HRP
  • Transparency Overlay Template
  • Wash Buffer Concentrate (25X)

 

Other Supplies Required

Reagents

Materials

  • Pipettes and pipette tips
  • Gloves
  • Plastic container with the capacity to hold 50 mL (for washing the arrays)
  • Plastic transparent sheet protector (trimmed to 10 cm x 12 cm and open on three sides)
  • Plastic wrap
  • Absorbent lab wipes (KimWipes® or equivalent)
  • Paper towels
  • X-ray film (Kodak BioMax™ Light-1) or equivalent
  • Flat-tipped tweezers

Equipment

  • Rocking platform shaker
  • Microcentrifuge
  • Autoradiography cassette
  • Film developer
  • Flatbed scanner with transparency adapter capable of transmission mode
  • Computer capable of running image analysis software and Microsoft Excel

Other Supplies Required for Cell Lysate Samples

  • Phosphate-Buffered Saline (PBS)
  • Lysis buffer (1% Igepal CA-630, 20 mM Tris-HCl (pH 8.0), 137 mM NaCl, 10% glycerol, 2 mM EDTA, 10 µg/mL Aprotinin, 10 µg/mL Leupeptin, and 10 µg/mL Pepstatin)

Other Supplies Required for Tissue Lysate Samples

  • PBS with protease inhibitors (10 µg/mL Aprotinin, 10 µg/mL Leupeptin, and 10 µg/mL Pepstatin)
  • Triton X-100

 

Procedure Overview

R&D Systems Protocol for Multiple Analyte Detection Using the Proteome Profiler Mouse XL Cytokine Array Kit, Panel A (Catalog # ARY028)

Add 2 mL of Array Buffer 6 to each well of the supplied 4-Well Multi-dish.

Add 2 mL of Array Buffer 6 to each well of the supplied 4-Well Multi-dish.

Place each array membrane in a separate well of the 4-Well Multi-dish.

Incubate for one hour on a rocking platform shaker.

Place each array membrane in a separate well of the 4-Well Multi-dish

Add 0.5 mL Array Buffer 4 to each sample.

Adjust volume of each sample to final volume of 1.5 mL with Array Buffer 6.

Add 0.5 mL Array Buffer 4 to each sample.

Replace the Array Buffer 6 in each well of the 4-Well Multi-dish with prepared samples.

Incubate overnight at 2 °C to 8 °C on a rocking platform.

Wash each array membrane 3 times with 1X Wash Buffer in a separate container.

Wash each well of the 4-Well Multi-dish with 1X Wash Buffer.

Add 0.5 mL Array Buffer 4 to each sample.
Add 30 µL of Detection Antibody Cocktail to 1.5 mL of Array Buffer 4/6 for each array.
Add 0.5 mL Array Buffer 4 to each sample.

Pipette 1.5 mL diluted Detection Antibody Cocktail into each well of the 4-Well Multi-dish.

Add 0.5 mL Array Buffer 4 to each sample.

Return each array membrane to the 4-Well Multi-dish containing diluted Detection Antibody Cocktail.

Incubate for one hour on a rocking platform shaker.

Wash each array membrane 3 times with 1X Wash Buffer in a separate container.

Wash each well of the 4-Well Multi-dish with 1X Wash Buffer.

Add 0.5 mL Array Buffer 4 to each sample.

Add 2 mL of diluted Streptavidin-HRP to each well of the 4-Well Multi-dish.

Place the array membrane in the diluted Streptavidin-HRP solution.

Incubate for 30 minutes on a rocking platform shaker.

Wash each array membrane 3 times with 1X Wash Buffer in a separate container.

Add 0.5 mL Array Buffer 4 to each sample.

Place the array membrane on a plastic sheet protector.

Pipette 1 mL of the prepared Chemi Reagent Mix evenly onto the membrane.

Cover the membrane with the top sheet of the plastic protector.

Incubate for 1 minute.

Blot off excess Chemi Reagent Mix.

Wrap the membrane and sheet protector in plastic wrap.

Place the wrapped array membrane in an autoradiography film cassette and expose to X-ray film.

Add 0.5 mL Array Buffer 4 to each sample.
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

  1. Metabolic origins of spatial organization in the tumor microenvironment
    Authors: C Carmona-Fo, M Deforet, L Akkari, CB Thompson, JA Joyce, JB Xavier
    Proc. Natl. Acad. Sci. U.S.A, 2017;0(0):. 2017
  2. Promyelocytic Leukemia Protein (PML) Controls Listeria monocytogenes Infection
    Authors: D Ribet, V Lallemand-, O Ferhi, MA Nahori, H Varet, H de Th?, P Cossart
    MBio, 2017;8(1):. 2017
  3. Astrocyte-derived interleukin-15 exacerbates ischemic brain injury via propagation of cellular immunity
    Proc. Natl. Acad. Sci. U.S.A, 2016;0(0):. 2016
Average Rating: 5 (Based on 4 reviews)

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 Proteome Profiler Mouse XL Cytokine Array ARY028
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Excellent
  Anonymous 05/23/2017
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 Proteome Profiler Mouse XL Cytokine Array ARY028
: Proteome Profiler Mouse XL Cytokine Array [ARY028]
 Proteome Profiler Mouse XL Cytokine Array ARY028
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Excellent
  Anonymous 11/28/2016
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 Proteome Profiler Mouse XL Cytokine Array ARY028
: Proteome Profiler Mouse XL Cytokine Array [ARY028]

Other Experimental Details

Other Experimental Details150 uL of mouse serum
  Excellent
  Anonymous 09/30/2016
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Other Experimental Details

Other Experimental DetailsThis is a great kit. It provides excellent reliability and a large number of cytokines, chemokines to screening in a single assay. I strongly recommend it.
 Proteome Profiler Mouse XL Cytokine Array ARY028
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Excellent
  Anonymous 08/26/2016
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 Proteome Profiler Mouse XL Cytokine Array ARY028
: Proteome Profiler Mouse XL Cytokine Array [ARY028]

Other Experimental Details

Other Experimental DetailsThe sample was cell culture supernatant collected from MEF.

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