FRS2 Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

FGFs (Fibroblast Growth Factors) bind to FGF receptors (FGFRs) monovalently, and FGF receptor dimerization and activation is mediated by multivalent interactions between heparin sulfate proteoglycans and FGF. Upon activation, receptor tyrosine kinases undergo rapid autophosphorylation on numerous tyrosine residues. Autophosphorylation sites located within the catalytic domain are crucial for stimulation of kinase activity, while autophosphorylation sites located in other regions are usually involved in the recruitment of cellular target proteins. Fibroblast growth factor or nerve growth factor (NGF) stimulation leads to tyrosine phosphorylation of the FGF Receptor Substrate 2 (FRS2) docking proteins. FRS2 is also designated SNT (Suc-1 associated neurotrophic factor target protein). Two structurally similar forms of FRS2 are: FRS2a (SNT-1) and FRS-2b (SNT-2, FRS3). FRS functions as a lipid-anchored docking protein that is tyrosine phosphorylated and recruited to FGFR upon FGF stimulation. FRS2 plays an important role in linking FGF and NGF with the Ras/MAPK signaling pathway, thus relaying information from the cell surface to the nucleus. FRS2 contains both a consensus myristylation sequence, involved in its recruitment to the cell membrane and a putative phosphotyrosine binding (PTB)domain in its amino-terminus. The interaction of FRS2 with FGFR1 occurs via the PTB domain of FRS2 binding to a 12-residue segment in the juxtamembrane region of FGFR1. FRS2a contains four binding sites for the adaptor protein Grb2 and two binding sites for the protein tyrosine phosphatase Shp2. FGF stimulation leads to phosphorylation of Shp2 on a tyrosine residue forming a complex with an additional molecule of Grb2. Grb2/Sos complexes are thus recruited directly and indirectly via Shp2 upon tyrosine phosphorylation of FRS2a in response to growth factor stimulation. It has been shown that FRS2 proteins are tyrosine phos phorylated in response to activation of the RET receptor, a tyrosine kinase that functions as the signal transducing receptor for the GDNF.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Array NBP2-09242

• FRS2 Antibodies
• FRS2 Lysates
• FRS2 Proteins