- PA1-662 - Invitrogen Antibodies DNM3 antibody

Submitted references The Expression of Dynamin 1, 2, and 3 in Human Hepatocellular Carcinoma and Patient Prognosis.

Tian M, Yang X, Li Y, Guo S
Medical science monitor : international medical journal of experimental and clinical research 2020 Jun 23;26:e923359

PICK1 regulates AMPA receptor endocytosis via direct interactions with AP2 α-appendage and dynamin.

Fiuza M, Rostosky CM, Parkinson GT, Bygrave AM, Halemani N, Baptista M, Milosevic I, Hanley JG
The Journal of cell biology 2017 Oct 2;216(10):3323-3338

Dynamin2 GTPase contributes to invadopodia formation in invasive bladder cancer cells.

Zhang Y, Nolan M, Yamada H, Watanabe M, Nasu Y, Takei K, Takeda T
Biochemical and biophysical research communications 2016 Nov 18;480(3):409-414

Stabilization of actin bundles by a dynamin 1/cortactin ring complex is necessary for 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 2013 Mar 6;33(10):4514-26

A missense mutation in a highly conserved alternate exon of dynamin-1 causes epilepsy in fitful mice.

Boumil RM, Letts VA, Roberts MC, Lenz C, Mahaffey CL, Zhang ZW, Moser T, Frankel WN
PLoS genetics 2010 Aug 5;6(8)

Beta-amyloid-induced dynamin 1 depletion in hippocampal neurons. A potential mechanism for early cognitive decline in Alzheimer disease.

Kelly BL, Vassar R, Ferreira A
The Journal of biological chemistry 2005 Sep 9;280(36):31746-53

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot of Dyn3 on rat liver extract using Product # PA1-662.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 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 (Product # PA1-662) 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 (Product # 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 (Product # 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4. PICK1 binds dynamin via BAR and GTPase domains and regulates dynamin polymerization. (A) Endogenous PICK1 interacts with endogenous dynamin 1, 2, and 3 in neurons. Extracts of cortical neurons were immunoprecipitated with PICK1 antibody or control IgG. Proteins were detected by Western blotting. Input is 5% of offered protein. Blots shown are representative of more than five independent experiments. (B) PICK1 interacts directly with dynamin. GST or GST-PICK1 was immobilized on glutathione agarose and incubated with purified HA-dynamin. Proteins were detected by Western blotting. Blots shown are representative of four independent experiments. (C) PICK1 interacts with the GTPase domain of dynamin. HEK293 cells were cotransfected with plasmids expressing Flag-PICK1 and GFP, GFP-dynamin 2, or dynamin domains as indicated. Cells were lysed and incubated with GFP-trap agarose. Blots shown are representative of five independent experiments. (D) Dynamin interacts with the PICK1 BAR domain. HEK cells were cotransfected with plasmids expressing HA-dynamin and GFP, GFP-PICK1, or truncations, as indicated in the top panel. Cells were lysed and incubated with GFP-trap agarose. Blots shown are representative of five independent experiments. (E) PICK1 promotes dynamin polymerization. His 6 -PICK1 at the concentrations indicated was incubated with HA-dynamin. Polymerized dynamin was pelleted by centrifugation, and protein in the supernatant and pellet was analyzed by Western bl

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4 Developmental expression patterns of dynamin-1 isoforms. (A) mRNA expression levels of Dnm1 isoforms during development in wildtype and mutant animals. The variant isoform region is amplified with common primers and the two transcripts are distinguished by a diagnostic Hph I restriction enzyme site specific for the b isoform. The two bands representing the Dnm1b transcript cDNA run as one band and lower on the gel than the Dnm1a transcript cDNA. Note that the 1a isoform becomes increasingly upregulated during development, while the b isoform is down regulated, however this ratio of a to b transcripts is skewed in the mutant animals. Below the gel is a schematic outlining the basis of the assay. Additional replicates are shown in Figure S2 . (B) Left , quantification of Dnm1b mRNA expression as assayed by PCR and visualized on agarose gels, relative to total Dnm1 mRNA in whole brain of E17.5, P0, and P14 wildtype and mutant brains. Data are expressed as the mean (+- SEM) relative proportion of Dnm1b to Dnm1 mRNA in the total cleaved PCR product. *P

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