Proteome Profiler Human XL Cytokine Array Kit

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, human milk and saliva.

Principle of the Array

The Proteome Profiler Human 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.

• Rectangular 4-Well Multi-dish
• 4 Human XL Cytokine Array nitrocellulose membranes spotted with 105 different antibodies to human cytokines
• Array Buffer 4
• Array Buffer 6
• Chemi Reagent 1
• Chemi Reagent 2
• Detection Antibody Cocktail, Human 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.

Stability and Storage

Store the unopened kit at 2 ^°C to 8 ^°C. Do not use past kit expiration date.

• Assays for Analytes represented in the Human XL Cytokine Array Kit
• Troubleshooting Guide

The Human XL Cytokine Array detects multiple cytokines, chemokines, growth factors and other soluble proteins in cell culture supernates. Cells were untreated or treated as indicated below. 500 µL of cell culture supernate was run on each array. Data shown are from a 10 minute exposure to X-ray film. Profiles of mean spot pixel density were created using a transmission-mode scanner and image analysis software.

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Figure 1A. HUVEC human umbilical vein endothelial cells were untreated or treated with 100 µg/mL recombinant human TNF-alpha (Catalog #210-TA) for 24 hours.

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Figure 1B. SW480 human colorectal adenocarcinoma cells were untreated or treated with 100 ng/mL of LPS for 24 hours.

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Figure 1C. THP-1 human acute monocytic leukemia cells untreated or treated with 1 µg/mL of recombinant human IFN-gamma (Catalog #285-IF) for 16 hours and 1 µ/mL of LPS for 8 hours.