72254 WideScreen® Phospho-EGFR Profiling Panel 9-Plex Assay Kit
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| Overview | This product has been discontinued. This product has been discontinued. We are currently offering a comparable kit, as a fixed panel, with the same analytes, with the exception of EGFR (Tyr1173, available in the MILLIPLEX MAP magnetic bead format (Cat. No. 48-613MAG). Please read the alternative product documentation carefully and contact technical service if you need additional information. ***Please Note: Although we are now EMD Millipore, our ordering systems are still separate. Please complete your order before linking to this alternative, as you will navigate away from this page and may lose other items in your basket. You must place a separate order with Millipore for this alternative. We apologize for this inconvenience.*** Bead-based flow cytometric xMAP® assays enable sensitive, precise detection of analytes within a sample. When directed toward proteins, such assays are essentially sandwich immunoassays on a bead. Samples are combined with fluorescently labeled microparticles (beads) covalently conjugated to a capture antibody. Analytes captured on the beads are identified with detection antibodies and a fluorescent label. A major hallmark of bead-based assays over traditional protein detection methods (such as ELISA) is the capacity for multiplexing, as bead-based assays allow simultaneous detection of multiple analytes in a small sample volume. Other advantages of xMAP® assays include flexibility, robustness, simple sample handling, and requirement of minimal sample volumes, making them an ideal platform for immunodetection of biomarkers and signaling proteins. Receptor Tyrosine Kinases (RTKs) are important regulators of numerous cell signaling pathways and have been implicated in various disease states. Protein phosphorylation of RTKs plays a critical role in the signaling cascades that regulate cell proliferation and development. The Epidermal Growth Factor Receptor (EGFR) family of RTKs plays a key role in propagating signals regulating cell proliferation, differentiation, motility, and apoptosis. EGFR is a 175 kDa RTK that gets activated by ligands like the epidermal growth factor (EGF). EGF binding induces EGFR autophosphorylation at specific tyrosine residues. These phosphorylation events promote the docking of several SH2 domain-containing adaptor proteins to EGFR that, in turn, activate downstream signaling pathways involved in regulating cell growth and proliferation, e.g., the Ras-mediated Mitogen Activated Protein Kinase (MAPK) pathway. Other downstream effectors are involved in regulating cell survival and apoptosis, such as the phosphatidylinositol 3-kinase (PI3K)/Akt pathway, and the signal transduction and activator of transcription (STAT) pathway. In addition to autophosphorylated tyrosine residues, the EGFR receptor contains other phosphorylation sites including phosphothreonine and phosphoserine residues that are involved in cross-talk with other signaling pathways and EGFR down-regulation. The WideScreen® Phospho-EGFR Profiling Panel 9-Plex Assay Kit allows for the individual detection of various phosphorylation sites on EGFR, which include: • pT654 • pT669 • PY845 • pY1045 • pS1047 • pY1068 • pY1086 • pY1173 • total EGFR (Epidermal Growth Factor Receptor) Depending on the stimulation conditions, individual phosphorylation sites of the EGFR receptor may be phosphorylated differentially in target cell lines. In the WideScreen® assay, cellular protein samples are prepared by gentle membrane extraction. Extracts are combined with dye-labeled Luminex® xMAP® beads covalently conjugated to capture antibodies specific to each phosphorylation site. After incubation and several washes, the beads are incubated with biotin-labeled detection antibodies. The resulting bead-immobilized immunosandwich is detected with streptavidin-phycoerythrin and acquired using a Luminex® xMAP® instrument. |
| The WideScreen® Phospho-EGFR Profiling Panel 9-Plex Assay Kit includes the entire set of reagents to assay one 96-well plate, including the Site-Specific Phospho-EGFR 9-plex, the premixed Site-Specific Phospho-EGFR Control Peptide mixture, the Cell Extraction Kit, and the WideScreen® Reagent Kit. The premix of various control peptides contain enough material to generate control data for 6 or 12 control wells in duplicate. Since these peptides do not mimic the phosphorylation site in the cellular context, these assays are not quantitative, and only allow for the qualitative comparison of site-specific EGFR phosphorylation in response to stimuli or inhibition. | |
| Catalogue Number | 72254 |
| Brand Family | Novagen® |
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Documentation
WideScreen® Phospho-EGFR Profiling Panel 9-Plex Assay Kit Analysenzertifikate
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| 72254 |
Broschüre
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| BeadPlex®Assays Product Compendium |
Technische Informationen
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| TB534 WideScreenﾙ BeadPlexﾙ Phospho-EGFR Profiling Panel 9-Plex Complete Assay Kit |
Anwendungsvorschriften
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| TB534 WideScreen® Phospho-EGFR Profiling Panel 9-Plex Assay Kit |