Proteome Profiler Phospho-Kinase Array Kit, BioAssay™ (Human)

Analyzing the phosphorylation profiles of kinases and their protein substrates is essential for understanding how cells recognize and respond to changes in their environment. The Human Proteome Profiler Phospho-Kinase Array Kit, BioAssay™ is a rapid, sensitive, and economical tool to simultaneously detect the relative levels of phosphorylation of 37 kinase phosphorylation sites and 2 related total proteins without performing numerous immunoprecipitations and Western blots. Each capture antibody was carefully selected using cell lysates prepared from cell lines known to express the target protein.||The human Proteome Profiler Phospho-Kinase Array Kit, BioAssay™ 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.||Key Benefits:|1. Detects phosphorylation of 37 human kinases and total amounts of 2 related proteins simultaneously.|2. Requires no specialized equipment.||Principle of the Assay:|Capture and control antibodies have been spotted in duplicate on nitrocellulose membranes. Cell lysates are diluted and incubated overnight with the Human Phospho-Kinase Array. The array is washed to remove unbound proteins followed by incubation with a cocktail of biotinylated detection antibodies. Streptavidin-HRP and chemiluminescent detection reagents are applied and a signal is produced at each capture spot corresponding to the amount of phosphorylated protein bound. Refer to the Appendix for a list and coordinates of analytes and controls.||Kit Components:|Human Phospho-Kinase Array: 8 nitrocellulose membranes (4 Part A, 4 Part B) each containing 39 different capture antibodies printed in duplicate.|Array Buffer 1: 21ml of a buffered protein base with preservatives.|Array Buffer 2 Concentrate, (5X) 12ml of a concentrated buffered protein base with preservatives.|Array Buffer 3: 21ml of a buffered protein base with|Lysis Buffer 6: 21ml of a denaturing buffered solution.|Wash Buffer Concentrate: 2 vials (21ml/vial) of a 25-fold concentrated solution of buffered surfactant with preservative. Note: May turn yellow over time|Detection Antibody Cocktail A, Human Phospho-Kinase Array: 1 vial of biotinylated antibody cocktail; lyophilized.|Detection Antibody Cocktail B, Human Phospho-Kinase Array|Streptavidin-HRP: 1x200ul|Chemi Reagent 1: 1xvial (2.5ml)|Chemi Reagent 2: 1xvial (2.5ml)|8-Well Multi-dish|Transparency Overlay Template||Storage and Stability:|Unused kit is stable for 3 months at 4°C. Once opened may be stored for up to 3 months at 4°C; return unused membranes to the foil pouch containing the dessicant pack and reseal along entire edge of the zip seal.