Proteome Profiler Human Phospho-MAPK Array Kit

A membrane-based antibody array for the parallel determination of the relative phosphorylation of human mitogen-activated protein kinases.

Validated for analyte detection in cell lysates.

Troubleshooting Guide

Key Benefits

• Detects phosphorylation of 26 human kinases simultaneously
• Requires no specialized equipment

View Data Examples
 

Principle of the Assay

The Proteome Profiler Human Phospho-MAPK Array Kit is a membrane-based sandwich immunoassay. Samples are mixed with a cocktail of biotinylated phospho-specific 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 phosphorylation in the bound analyte.

Why Use an Antibody Array to Detect Receptor Phosphorylation?

Determining the phosphorylation of multiple kinases 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 phosphorylations in a single sample can be cost-effective and also save time and sample.

• 4-Array Membranes
• 4-Well Multi-dish
• Lysis Buffer
• Array Buffers
• Wash Buffer
• Detection Antibody Cocktail
• Streptavidin-HRP
• Chemiluminescent Detection Reagents
• Transparency Overlay Template
• Detailed product datasheet with protocol

For a complete list of the kit contents and necessary materials, please see the Materials Provided/Other Supplies Required sections of the product datasheet.

Assays for analytes represented in the Human Phospho-MAPK Array Kit

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

Figure 1.HeLa human cervical epithelial carcinoma cells were either untreated or exposed to 150 J/m2 of UV light followed by a 30 minute recovery period before lysis. Data shown are from a 2 minute exposure to X-ray film.

Figure 2. : MCF-7 human breast cancer cells were either untreated or treated with 100 ng/mL of rhIGF-I (R&D Systems, Catalog # 291-G1) for 1 hour. Cells for rhIGF-I treatment either received a 1 hour pre-treatment with the PI3K inhibitor, LY294002, or received no pre-treatment. Data shown are from a 6 minute exposure to X-ray film.

BG01V human embryonic stem cells are licensed from ViaCyte, Inc.

Quick Spots image analysis software

Protein kinases are the largest class of enzymes in the human genome. These enzymes regulate almost all cellular processes by adding phosphate groups to proteins, thereby modifying the activity, localization, and overall function of their targets. Consequently, abnormal activity of kinases underlies many diseases including cancer.