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Pathway Focus: The Integrin Pathway
Quantify Signaling Proteins Associated With Integrin Binding—An Easy and Sensitive Way to Measure Proteins in Cell Lysates
Looking for an easy way to measure focal adhesion kinase (FAK) and β-catenin proteins? Use phosphoELISA™ kits to quickly measure signaling proteins in cell lysates using a convenient 96-well format. Just process your samples as you would for western blotting, add them to an ELISA plate, run the assay, and get quantitative results in 4 hours.
Integrins play an important role in cell attachment to other cells or to the extracellular matrix as well as signal transduction. FAK is a non-receptor protein tyrosine kinase that localizes to focal adhesions. In response to integrin engagement, FAK is autophosphorylated at tyrosine residue 397. This autophosphorylation event creates high-affinity binding sites for the SH2 domains of several important signaling proteins. FAK is under investigation in several areas of research, including cell adhesion and migration studies, studies of migration in response to growth factors, differentiation, cell cycle, apoptosis, and cancer studies.
|FAK is overexpressed in breast, colon, and thyroid cancers, possibly contributing to some features of the malignant phenotype (i.e., increased migration and invasion of surrounding stroma).|
β-Catenin is part of a complex of proteins that constitute adherens junctions. β-Catenin also anchors the actin cytoskeleton and may be responsible for transmitting the contact inhibition signal that causes cells to stop dividing once the epithelial sheet is complete. Recent evidence suggests that β-catenin plays an important role in various aspects of liver biology, including liver development, liver regeneration following partial hepatectomy, HGF-induced hepatomegaly, and pathogenesis of liver cancer.
We offer phosphoELISA™ kits for both phosphorylation site–specific proteins and total proteins to elucidate specific target proteins of this cell signaling pathway.
|Figure 1. Detection of FAK using Invitrogen™ ELISA kits. 3T3L1 cells were treated with 1 mM sodium orthovanadate for 5 hr, and untreated 3T3L1 cells were used as control. Cell extracts were prepared in 0.1% SDS extraction buffer and analyzed using the FAK [pY397] ELISA and FAK (Total) ELISA. (A) The FAK [pY397] detected phosphorylated FAK recombinant protein and phosphorylated FAK in orthovanadate-treated 3T3L1 cells, but not the nonphosphorylated FAK recombinant protein or nonphosphorylated FAK in untreated 3T3L1 cells. (B) In contrast, the FAK (Total) ELISA kit detected both phosphorylated and nonphosphorylated FAK recombinant protein and FAK in orthovanadate-treated cells and untreated control.|
|Figure 2. Detection of β-catenin using Invitrogen™ ELISA kits. The data show that the β-catenin ELISA kit detects β-catenin protein in cell lysates from human GTL16, Jurkat, HeLa, MCF-7, and mouse NIH3T3 cells. The levels of β-catenin protein detected with this ELISA kit are consistent with results obtained by western blot analysis.|
|FAK [PY397] ELISA KIT||96 tests||KHO0441|
|FAK (Total) ELISA KIT||96 tests||KHO0431|
|HU B-catenin (Total) ELISA KIT||96 tests||KHO1211|
|Integrins are important in transmitting signals from the extracellular matrix (ECM) to cells. The ECM is involved in movement of cells during wound healing and embryonic development, and also relays important information to the cells. Two key ligands found in the ECM are fibronectin and vitronectin. These glycoproteins bind to integrins at the surface of the cell and play a role in a variety of biological processes, including cell adhesion, growth, and migration.||We offer natural human fibronectin and natural human vitronectin purified from human plasma for use in cell culture applications.|
|Natural Human Vitronectin||100 µg||PHE0011|| |
|Natural Human Fibronectin||1 mg||PHE0023|