Immunofluorescence analysis of Dynamin III was performed using 70% confluent log phase SH-SY5Y cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Dynamin III Rabbit Polyclonal Antibody (PA1-662) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A27034) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic Peptide: C P(623) D K S F T E N D E N G Q A E N F(639)|
|Purification||Ammonium sulfate precipitation|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:1,000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 3 publications below|
PA1-662 detects dynamin III (Dyn3) from rat, mouse, and human tissues and cells.
PA1-662 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~100 kDa protein representing Dyn3 from HeLa cell lysate.
PA1-662 immunizing peptide corresponds to amino acid residues 623-639 from rat Dyn3. PA1-662 immunizing peptide (Cat. # PEP-057) is available for use in neutralization and control experiments.
The dynamins are a family of 100 kDa GTPases transcribed from at least three separate genes. At least four mRNA splice variants for each dynamin have been described. Dynamins contain several conserved regions including the conserved, amino-terminal GTPase domain, a centrally located membrane-binding plekstrin homology domain (PHD), and a coiled-coil region located in front of a proline-rich domain (PRD). The PRD is thought to mediate interactions between dynamin and numerous other cellular proteins. Dynamin I is expressed exclusively in neurons, dynamin II is ubiquitously expressed, and dynamin III is thought to be restricted to expression in the brain, testis, heart, and lung. The dynamins participate in the cellular process of clathrin-mediated and fluid-phase endocytosis.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Stabilization of actin bundles by a dynamin 1/cortactin ring complex is necessary for growth cone filopodia.
PA1-662 was used in western blot to demonstrate the role of a dynamin 1/cortactin ring complex for the maintenance of growth cone filopodia
|Yamada H,Abe T,Satoh A,Okazaki N,Tago S,Kobayashi K,Yoshida Y,Oda Y,Watanabe M,Tomizawa K,Matsui H,Takei K||The Journal of neuroscience : the official journal of the Society for Neuroscience (33:4514)||2013|
A missense mutation in a highly conserved alternate exon of dynamin-1 causes epilepsy in fitful mice.
PA1-662 was used in western blot to study the role of a missense mutation in dynamin-1 in the pathogenesis of epilepsy in fitful mice
|Boumil RM,Letts VA,Roberts MC,Lenz C,Mahaffey CL,Zhang ZW,Moser T,Frankel WN||PLoS genetics (6:null)||2010|
Beta-amyloid-induced dynamin 1 depletion in hippocampal neurons. A potential mechanism for early cognitive decline in Alzheimer disease.
PA1-662 was used in western blot to demonstrate a potential mechanism for early cognitive decline in Alzheimer disease.
|Kelly BL,Vassar R,Ferreira A||The Journal of biological chemistry (280:31746)||2005|