Summary for Proteome Profiler Human Cytokine Array Kit
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, plasma, saliva, urine, and human milk.
Detects 36 human cytokines, chemokines, and acute phase proteins simultaneously
Requires no specialized equipment
Compatible with LI-COR* and chemiluminescence detection
*If using a LI-COR, additional reagents and protocol modifications are required. Refer to protocol.
Principle of the Assay
The Proteome Profiler Human Cytokine Array Kit is a membrane-based sandwich immunoassay. Samples are mixed with a cocktail of biotinylated detection antibodies (Step 1) and then incubated with the array membrane which is spotted in duplicate with capture antibodies to specific target proteins (Step 2). Captured proteins are 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
Rectangular 4-Well Multi-dish
Human Cytokine Array, Panel A nitrocellulose membranes spotted with 36 different antibodies to human cytokines, chemokines, and acute phase proteins
Array Buffer 4
Array Buffer 5
Chemi Reagent 1
Chemi Reagent 2
Detection Antibody Cocktail, Human 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 acute phase proteins in a single sample.
C5a
IL-4
IL-27
CD40 Ligand
IL-5
IL-32 alpha
G-CSF
IL-6
CXCL10/IP-10
GM-CSF
IL-8
CXCL11/I-TAC
CXCL1/GRO alpha
IL-10
CCL2/MCP-1
CCL1/I-309
IL-12 p70
MIF
ICAM-1
IL-13
MIP-1 alpha/MIP-1 beta
IFN-gamma
IL-16
CCL5/RANTES
IL-1 alpha
IL-17
CXCL12/SDF-1
IL-1 beta
IL-17E
Serpin E1/PAI-1
IL-1ra
IL-18
TNF-alpha
IL-2
IL-21
TREM-1
Assays for Analytes represented in the Human Cytokine Array Kit
Product Specifications for Proteome Profiler Human Cytokine Array Kit
Species
Human
Source
N/A
Scientific Data Examples for Proteome Profiler Human Cytokine Array Kit
The Human Cytokine Array detects multiple cytokines, chemokines, growth factors and other soluble proteins in cell culture supernates.
The amount of cell culture supernate used on each array and the duration of exposure to X-ray film is indicated below. Profiles of mean spot pixel density were created using a transmission-mode scanner and image analysis software. 1A: THP-1 human acute monocytic leukemia cells were untreated or treated with 1 µg/mL LPS for 4 hours, then 5mM Nigericin (Tocris, Catalog # 4312) for 1 hour (500 µL of cell culture supernate, 10 minute exposure). 1B: SW480 human colorectal adenocarcinoma cells were untreated or treated with 100 ng/mL LPS for 24 hours (500 µL of cell culture supernate, 10 minute exposure).
The Human Cytokine Array detects multiple cytokines, chemokines, growth factors and other soluble proteins in cell lysates.
The amount of cell lysate used on each array and the duration of exposure to X-ray film is indicated below. Profiles of mean spot pixel density were created using a transmission-mode scanner and image analysis software. 2A: SW1353 human chondrosarcoma cells were untreated or treated with 5 ng/mL recombinant human IL-1 alpha (R&D Systems, Catalog # 200-LA) and 10 ng/mL recombinant human OSM (R&D Systems, Catalog # 295-OM) for 24 hours (200 µg of cell lysate, 5 minute exposure). 2B: THP-1 human acute monocytic leukemia cells were untreated or treated with 1 µg/mL recombinant human IFN-gamma (R&D Systems, Catalog # 285-IF) for 8 hours, then 1 µg/mL LPS for 16 hours (200 µg of cell lysate, 5 minute exposure).
The Human Cytokine Array detects multiple cytokines, chemokines, growth factors and other soluble proteins in tissue lysate samples.
200 µg of tissue lysate was run on each array. Data shown are from a 5 minute exposure to X-ray film.