EDG 6 Blocking Peptide (Endothelial Cell Differentiation Gene 6), Purified

Endothelial Cell Differentiation Gene-6, or EDG-6, encodes a 45kD protein belonging to a family of G-Protein Coupled Receptors (GPCR's), which bind lysophospholipids. To date, 8 members of the EDG receptor family have been identified. These receptors are suggested to have a role in modulation of growth-related effects including cell proliferation and differentiation as well as protective, anti-apoptotic effects. EDG receptor expression is developmentally regulated and the expression of the 8 members of this family of proteins appears to be tissue specific. EDG receptors couple to G-proteins to signal through Ras and MAPK pathways and other protein tyrosine kinase pathways. EDG-6 is also known as S1P4. The ligand for EDG-6 is sphingosine-1-phosphate. EDG-6 is expressed by lymphoid and hematopoietic tissues and also by lung tissue. The biological function of EDG-6 is currently under investigation. Sphingosine-1-phosphate (S1P) exerts its activity by binding to five distinct G-protein-coupled receptors, S1P1/EDG-1, S1P2/EDG-5, S1P3/EDG-3, S1P4/EDG-6, and S1P5/EDG-8. S1P3 couples to Gi/o-ERK, Gq-PLC, and G12/13-Rho axes to mediate S1P-induced cell proliferation, survival, migration, and related signaling events. S1P3 is widely expressed in various tissues, suggesting diverse physiological functions of this receptor. The human and murine S1P3 have 378 amino acids with an estimated molecular weight of 42kD. Glycosylation at the N-terminal extracellular domain may cause the protein to migrate at different positions in SDS-PAGE. Cayman's S1P3 polyclonal antibody detects the receptor at 47kD using samples from human cerebral cortex. Liver and heart tissues display 2-3 bands between 40-50kD, possibly due to different degree of glycosylation. , Applications: Blocking peptide for use with EDG-6 antibody to demonstrate the specificity of this antibody. Suggested Working Dilutions: For use as a control in Western blotting, it is recommended that the peptide be used at a 50 fold molar excess over the antibody. The antibody, used neat, should be pre-incubated with the peptide for 20 minutes at 37°C. Following this pre-incubation step, the antibody should be diluted to the appropriate concentration for use in Western blotting and applied to the membrane. The antibody pre-incubated with an equivalent volume of phosphate buffered saline only may also be included as a control. The optimal concentration should be determined for each specific application. , Storage and Stability: , Lyophilized powder may be stored at 4°C for short-term only. Stable for 12 months at -20°C. Reconstitute to nominal volume (see reconstitution instructions for peptides) and store at -20°C. For maximum recovery of product, centrifuge the original vial prior to removing the cap. Further dilutions can be made in assay buffer.