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Product Details

Proteome Profiler Mouse Cytokine Array Kit, Panel A Summary

Kit Summary

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

Key Benefits

l Detects 40 mouse cytokines, chemokines, and acute phase proteins simultaneously

l Requires no specialized equipment

l 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 Mouse Cytokine Array Kit, Panel A 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

l Rectangular 4-Well Multi-dish

l 4 Mouse Cytokine Array, Panel A nitrocellulose membranes spotted with 40 different antibodies to mouse cytokines, chemokines, and acute phase proteins

l Array Buffer 4

l Array Buffer 6

l Chemi Reagent 1

l Chemi Reagent 2

l Detection Antibody Cocktail, Mouse Cytokine Array, Panel A

l Streptavidin-HRP

l Transparency Overlay Template

l 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.

Supporting data and figures

The Mouse Cytokine Array, Panel A utilizes capture antibodies spotted onto a nitrocellulose membrane to allow high-throughput multi-analyte profiling of 40 cytokines, chemokines, and more in a single sample.

Mouse splenocytes treated with 100 ng/mL LPS for 20 hours were mixed with a cocktail of biotinylated detection antibodies, and then incubated with the Mouse Cytokine Array. The array was then incubated with streptavidin-horseradish peroxidase followed by chemiluminescent detection. After detection, the array data were quantitated to generate a protein profile (histogram). The table shows the analytes detected and their location on the membrane.

The Mouse Cytokine Array detects multiple analytes in cell culture lysates and supernates.

(A. and B.) Lysates of RAW264.7 mouse monocyte/macrophage cell line untreated or treated with 100 ng/mL LPS for 24 hours. 100 μg of lysate was used for each array shown. (C. and D.) Supernates of RAW264.7 mouse monocyte/macrophage cell line untreated or treated with 100 ng/mL LPS for 24 hours. 100 μL of supernate was used for each array shown. Array images were collected and analyzed using the LI-COR Odyssey Infrared Imaging System.

The Mouse Cytokine Array detects multiple analytes in mouse serum and tissue lysates.

(A and B.) Samples of mouse serum from two different vendors. 200 μL of serum was used for each array shown. (C. and D.) Lysates of mouse lung tissue and mouse stomach tissue. 200 μg of lysate was used for each array shown. Array images were collected and analyzed using the LI-COR Odyssey Infrared Imaging System.

Specifications