Immunofluorescence analysis of Dynamin II was performed using 70% confluent log phase HeLa 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 II Rabbit Polyclonal Antibody (PA1-661) at 2 µg/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 Alexa Fluor® 555 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||Rat, Non-human primate, Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic Peptide: C S(760) P T P Q R R P V S S I H P P G R P P A(779)|
|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 *|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Western Blot (WB)||2-4 µg/ml|
* 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.
PA1-661 detects dynamin II (Dyn2) from rat, mouse, and human tissues and cells.
PA1-661 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~100 kDa protein representing Dyn2 from HeLa cell lysate.
PA1-661 immunizing peptide corresponds to amino acid residues 760-779 from human Dyn2. This sequence is 95% conserved in mouse and rat Dyn2. PA1-661 immunizing peptide (Cat. # PEP-056) 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.
Uptake of Helicobacter pylori vesicles is facilitated by clathrin-dependent and clathrin-independent endocytic pathways.
PA1-661 was used in western blot to study the role of clathrin-dependent and clathrin-independent routes in the uptake of Helicobacter pylori vesicles by gastric epithelial cells
|Olofsson A,Nygård Skalman L,Obi I,Lundmark R,Arnqvist A||mBio (5:e00979)||2014|
Protein partners of dynamin-1 in the retina.
PA1-661 was used in immunohistochemistry and western blot to identify protein binding partners of retinal dynamin-1
|Grossman GH,Ebke LA,Beight CD,Jang GF,Crabb JW,Hagstrom SA||Visual neuroscience (30:129)||2013|
Induction of the alternative NF-¿B pathway by lymphotoxin ¿ß (LT¿ß) relies on internalization of LTß receptor.
PA1-661 was used in western blot to study the mechanism by which the alternative NF-kappaB pathway is induced by lymphotoxin alphabeta
|Ganeff C,Remouchamps C,Boutaffala L,Benezech C,Galopin G,Vandepaer S,Bouillenne F,Ormenese S,Chariot A,Schneider P,Caamaño J,Piette J,Dejardin E||Molecular and cellular biology (31:4319)||2011|
Dynamin 2 is required for the enhancement of HIV-1 infectivity by Nef.
PA1-661 was used in western blot to study to the role of dynamin 2 in HIV-1 infectivity.
|Pizzato M,Helander A,Popova E,Calistri A,Zamborlini A,Palù G,Göttlinger HG||Proceedings of the National Academy of Sciences of the United States of America (104:6812)||2007|
Beta-amyloid-induced dynamin 1 depletion in hippocampal neurons. A potential mechanism for early cognitive decline in Alzheimer disease.
PA1-661 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|
Analysis of clathrin-mediated endocytosis of epidermal growth factor receptor by RNA interference.
PA1-661 was used in western blot to assess the importance of clathrin, AP-2, and other endocytic proteins for EGFR endocytosis.
|Huang F,Khvorova A,Marshall W,Sorkin A||The Journal of biological chemistry (279:16657)||2004|
|Non-human primate||Not Cited||
Detergent-resistant membrane microdomains facilitate Ib oligomer formation and biological activity of Clostridium perfringens iota-toxin.
PA1-661 was used in western blot to locate iota b (Ib) oligomers in detergent-resistant membrane microdomains (DRMs) extracted from Vero cells
|Hale ML,Marvaud JC,Popoff MR,Stiles BG||Infection and immunity (72:2186)||2004|
Aquaporin-2 localization in clathrin-coated pits: inhibition of endocytosis by dominant-negative dynamin.
PA1-661 was used in immunocytochemistry to investigate the aquaporin-2 localization in clathrin-coated pits.
|Sun TX,Van Hoek A,Huang Y,Bouley R,McLaughlin M,Brown D||American journal of physiology. Renal physiology (282:F998)||2002|