The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 - 5 µg/ml. Predicted molecular weight: 82 kDa.
Use a concentration of 1 µg/ml.
Use at an assay dependent concentration.
Use 1µg for 106 cells. ab170192-Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
機能Functions in mitochondrial and peroxisomal division. Mediates membrane fission through oligomerization into ring-like structures which wrap around the scission site to constict and sever the mitochondrial membrane through a GTP hydrolysis-dependent mechanism. Required for normal brain development. Facilitates developmentally-regulated apoptosis during neural tube development. Required for a normal rate of cytochrome c release and caspase activation during apoptosis. Also required for mitochondrial fission during mitosis. May be involved in vesicle transport. Isoform 1 and isoform 4 inhibit peroxisomal division when overexpressed.
組織特異性Ubiquitously expressed with highest levels found in skeletal muscles, heart, kidney and brain. Isoform 1 is brain-specific. Isoform 2 and isoform 3 are predominantly expressed in testis and skeletal muscles respectively. Isoform 4 is weakly expressed in brain, heart and kidney. Isoform 5 is dominantly expressed in liver, heart and kidney. Isoform 6 is expressed in neurons.
関連疾患Note=May be associated with Alzheimer disease through beta-amyloid-induced increased S-nitrosylation of DNM1L, which triggers, directly or indirectly, excessive mitochondrial fission, synaptic loss and neuronal damage.
配列類似性Belongs to the dynamin family. Contains 1 GED domain.
ドメインThe GED domain folds back to interact, in cis, with the GTP-binding domain and middle domain, and interacts, in trans, with the GED domains of other DNM1L molecules, and is thus critical for activating GTPase activity and for DNM1L dimerization.
翻訳後修飾Phosphorylation/dephosphorylation events on two sites near the GED domain regulate mitochondrial fission. Phosphorylation on Ser-637 inhibits mitochondrial fissin probably through preventing intramolecular interaction. Dephosphorylated on this site by PPP3CA which promotes mitochondrial fission. Phosphorylation on Ser-616 also promotes mitochondrial fission. Sumoylated on various lysine residues within the B domain. Desumoylated by SENP5 during G2/M transition of mitosis. Appears to be linked to its catalytic activity. S-nitrosylation increases DNM1L dimerization, mitochondrial fission and causes neuronal damage. Ubiquitination by MARCH5 affects mitochondrial morphology.
細胞内局在Cytoplasm > cytosol. Golgi apparatus. Endomembrane system. Mainly cytosolic. Translocated to the mitochondrial membrane through interaction with FIS1. Colocalized with MARCH5 at mitochondrial membrane. Localizes to mitochondria at sites of division. Associated with peroxisomal membranes, partly recruited there by PEX11B. May also be associated with endoplasmic reticulum tubules and cytoplasmic vesicles and found to be perinuclear. In some cell types, localizes to the Golgi complex.
DNM1L was immunoprecipitated using 0.5mg Hek293 whole cell extract, 10µg of Mouse monoclonal to DNM1L and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, Hek293 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation. Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab56788. Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution. Band: 90kDa; DNM1L.
Western blot - Anti-DRP1 antibody (ab56788)Image from PLoS Pathog. 2010 Jul; 6(7): e1001012. Fig 9E, doi: 10.1371/journal.ppat.1001012
HeLa cells were transfected with N.C. siRNA or hDRP1-targeted siRNA (#1–#3) for 72 h, and the knockdown of endogenous DRP1 was analyzed by Western blotting using anti-DRP1 antibody ab56788.
Immunocytochemistry/ Immunofluorescence - Anti-DNM1L antibody (ab56788)This image is courtesy of an anonymous Abreview
ab56788 staining overexpressed Human DNM1L in the HEK293 cell line by ICC/IF(Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde, permeabilized with Triton X-100 1% and blocked with 5% Serum for 60 minutes at 21°C. Samples were incubated with primary antibody (1/200 in 5% Horse serum) for 2 hours. An Alexa Fluor®488-conjugated Goat anti-mouse IgG polyclonal(1/400) was used as the secondary antibody. Results showed that overexpressed hDNM1L co-localizes with mitochondria.
DNM1L antibody (ab56788) used in immunohistochemistry at 1ug/ml on formalin fixed and paraffin embedded human leiomyosarcoma tissue.
Flow Cytometry - DNM1L antibody (ab56788)
Overlay histogram showing HEK293 cells stained with ab56788 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab56788, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HEK293 cells fixed with 100% methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
Anti-DRP1 antibody (ab56788) 使用論文
This product has been referenced in:
Pant M et al. Metabolic dysfunction and altered mitochondrial dynamics in the utrophin-dystrophin deficient mouse model of duchenne muscular dystrophy. PLoS One10:e0123875 (2015).
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