Phospho-Dynamin (S774) Antibody Summary
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Detection of Phospho-Dynamin (S774) by Western Blot. Western blot of rat brain (hippocampus) tissue lysate stimulated with forskolin showing specific immunolabeling of the ~95 kDa dynamin phosphorylated at S774 (Control). The phosphospecificity of this labeling is dem-onstrated by treatment with 1200 U of lambda Phosphatase ( lambda -PPase) for 30 minutes before being exposed to the Anti-Phospho-Dynamin (S774). The immunolabeling is completely eliminated by treatment with lambda -PPase.
Preparation and Storage
Rat Dynamin (or Dynamin 1; also known as dephosphin) is a 95-100 kDa GTPase that belongs to the “classic dynamin” group of the dynamin superfamily. It is 864 amino acids (aa) in length, and contains three N-terminal GTP-binding elements (aa 38-208), one pleckstrin-homology domain (aa 519-625), a GED (GTPase effector domain) (aa 645-745), and a C-terminal SH3-binding proline-rich region (aa 753-864). At least four alternate splice forms are known, one that shows a 47 aa substitution between aa 398-445, and three that generate unique C-termini over the last 20 amino acids. Dynamin is found constitutively phosphorylated and inactive in the cytoplasm just under synaptic membranes. Upon stimulation, calcineurin dephosphorylates, and nitric oxide nitrosylates, Dynamin. Dephosphorylated and nitrosylated Dynamin now interacts with the cell membrane, generating endocytic synaptic vesicles. The process terminates when Cdk5 rephosphorylates Dynamin on S744 and S778.
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