740 Y-P (PDGFR 740Y-P)

The PDGFR 740Y-P peptide stimulates a mitogenic response in muscle cells. The ability of the 740Y-P peptide to stimulate mitogenesis is highly specific and not a general feature of a cell permeable SH2 domain binding peptides^[2].

740Y-P is as effective as a growth factor (FGF2) at promoting neuronal cell survival via the established PI 3-kinase-Akt survival cascade. Stimulation of neuronal cell survival by the PDGFR740Y-P peptide is dose dependent and does not require insulin^[1].

740 Y-P decreases the degree of ROS levels in Aβ(25-32) treated hippocampal tissues and increases the extent of AKT and PI3K phosphorylation in Alzheimer’s disease rat model.

• Cell lines

  NIH 3T3 cells

• Concentrations

  50 μg/ml

• Incubation Time

  2 h

• Method

  NIH 3T3 cells (2×10⁶) are incubated in suspension at 37°C in Ca2+ and Mn2+ free HBSS, 10% FCS containing either 50 μg/ml of 740Y-P peptide or an equal volume of PBS as a control. After 2h cells are centrifuged, washed and trypsinised to degrade non-internalised peptide. Cells are then resuspended in lysis buffer (50 mM Tris pH 7.4, 150 mM NaCl, 10% Glycerol, 2% NP40, 0.25% deoxycholate, 1 mM EDTA, 1 mM Vanadate, Protease inhibitors "complete" cocktail from Boehringer-Mannheim) for 1 h at 4°C. Lysates are clarified by centrifugation at 1.4×10⁴g for 5 min and the supernatants are incubated for 1h with streptavidin-agarose beads. Beads are subsequently washed three times in lysis buffer and finally boiled in SDS sample buffer and resolved by SDS-PAGE on a 12% gel, transferred to nitrocellulose and immunoblotted with the p85 monoclonal antibody.