Detects phosphorylation of 37 human kinases and total amounts of 2 related proteins simultaneously
Requires no specialized equipment
Principle of the Assay
The Proteome Profiler Human Phospho-Kinase 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 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.
Each Human Phospho-Kinase Antibody Array Kit contains most of the necessary components for a ready-to-run assay without the need for specialized equipment.
8 Array Membranes (4 of membrane A and 4 of membrane B)
Detection Antibody Cocktail A
Detection Antibody Cocktail B
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.
Stability and Storage
Store the unopened kit at 2°C to 8°C. Do not use past kit expiration date.
Simultaneously detect the relative phosphorylation of these proteins in a single sample.
Akt 1/2/3 (S473)
Akt 1/2/3 (T308)
JNK 1/2/3 (T183/Y185, T221/Y223)
p38 alpha (T180/Y182)
EGF R (Y1086)
ERK1/2 (T202/Y204, T185/Y187)
P70 S6 Kinase (T389)
GSK-3 alpha/beta (S21/S9)
p70 S6 Kinase (T421/S424)
GSK-3 beta (S9)
PDGF R beta (Y751)
PLC gamma-1 (Y783)
Assays for analytes represented in the Human Phospho-Kinase Array Kit
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.
Product Specifications for Proteome Profiler Human Phospho-Kinase Array Kit
Scientific Data Examples for Proteome Profiler Human Phospho-Kinase Array Kit
Detection of Human Protein Kinase Phosphorylation in A549 Cell Line.
A549 human lung adenocarcinoma cells were either left untreated or exposed to 50 J/m2 of UV light followed by a 24 hour recovery before lysis.
Detection of Human Protein Kinase Phosphorylation in Daudi Cell Line.
Daudi human B cell lymphoma cells were either left untreated or treated with 500 U/ml recombinant human IFN-alpha 2a (R&D Systems®, Catalog # 11100-1) for 5 minutes.
Detection of Human Protein Kinase Phosphorylation in Jurkat Cell Line.
Jurkat human acute T cell leukemia cells were either left untreated or treated with 100 ng/mL PMA and 0.5 mM Ionomycin for 20 minutes.
Detection of Human Protein Kinase Phosphorylation in MCF-7 Cell Line.
MCF-7 human breast cancer cells were either left untreated or treated with 100 ng/mL of recombinant human IGF-1 (R&D Systems®, Catalog # 291-G1) for 1 hour.
Preparation & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
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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