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

Produktübersicht

• Artikelname: Proteome Profiler Phospho-Immunoreceptor Array Kit, BioAssay(TM) (Human)
• Artikelnummer: USB-052380
• Hersteller Artikelnummer: 052380
• Alternativnummer: USB-052380-1
• Hersteller: US Biological
• Kategorie: Kits/Assays

Produktbeschreibung

The tyrosine phosphorylation of immunoreceptors and adaptor signaling molecules on ITAMs (immunoreceptor tyrosine-based activation motifs) and ITIMs (immunoreceptor tyrosine-based inhibitory motifs) plays an important role in the control of cellular activation in the immune system. The Human Phospho-Immunoreceptor Array is a rapid, sensitive, and economical tool used to detect changes in phosphorylation between samples. Protein array technology allows the screening of fifty-nine different immunoreceptors without numerous individual immunoprecipitations and Western blots. The non-denaturing conditions used in the methods recommended for this array maintain the non-covalent association of activating receptors with phosphorylated ITAM-containing transmembrane signaling adaptors. The efficacy of each capture antibody was confirmed using lysate samples prepared from treated cell lines known to express the immunoreceptors. Recombinant immunoreceptors were used to validate capture antibodies by competition with lysates prepared from activated cell lines. The Proteome Profiler Human Phospho-Immunoreceptor 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). Tyrosine phosphorylation of the captured proteins is detected with an HRP-conjugated pan phospho-tyrosine antibody (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 59 human ITAM/ITIM-associated immunoreceptors 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 with the Human Phospho-Immunoreceptor Array. After binding the extracellular domain of both phosphorylated and unphosphorylated immunoreceptors, unbound material is washed away. A pan anti-phospho-tyrosine antibody conjugated to horseradish peroxidase (HRP) is then used to detect phosphorylated tyrosines on activated receptors by chemiluminescence. Kit Components: Human Phospho-Immunoreceptor Array, 4 membranes Array Buffer 1: 1 vial (21ml) Array Buffer 2 Concentrate, 5X: 1 vial (21ml) Lysis Buffer 17: 1 vial (21ml) Wash Buffer Concentrate, 25X: 2 vials (21ml/vial) Chemi Reagent 1: 1 vial (2.5 mL) Chemi Reagent 2: 1 vial (2.5 mL) Anti-Phospho-Tyrosine-HRP Detection Antibody: 1 vial (50uL) 4-Well Rectangular Multi-dish: 1 dish Transparency Overlay Template Storage and Stability: Unused kit is stable for 3 months at 4C. Once opened may be stored for up to 3 months at 4C, return unused membranes to the foil pouch containing the dessicant pack and reseal along entire edge of the zip seal.