Proteome Profiler Human Cytokine Array 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.
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
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.
MIP-1 alpha/MIP-1 beta
Assays for Analytes represented in the Human Cytokine Array Kit
Proteome Profiler Human Cytokine Array Kit Scientific Data Examples
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 (50 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.
Tested condition media of MSCs from the umbilical cord co-cultured with NK cells. The goal of this experiments was to detect chages in inflammatory factors in the system when MSCs were cultured with NK cells. The lack of detected secreted proteins is promising because this suggests that MSCs do not up-regulate large inflammatory proteins following immune cell activation. Left image : MSC cultured in RPMI media (Control)Right image: MSC co-cultured with NK cells in same RPMI media
Will the Array identification number stamped on the Array membrane interfere with detection if it is not cut-off before the membrane is blocked?
The dye used for printing the Array identification number on the membranes will fluoresce and interfere with the LI-COR detection. It is critical that the number is cut off before beginning the experiment.
Which Brands are Currently Available on bio‑techne.com? R&D Systems, Tocris Bioscience and ProteinSimple branded products are available to purchase through bio‑techne.com. ProteinSimple branded instruments are available to quote. ACD branded products will be available on bio‑techne.com in the near future. Novus Biologicals branded products are not currently available on bio‑techne.com and can be found at novusbio.com.
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