Recombinant
RabMAb

Anti-DRP1 antibody [EPR19274] (ab184247)

Overview

  • Product name
    Anti-DRP1 antibody [EPR19274]
    See all DRP1 primary antibodies
  • Description
    Rabbit monoclonal [EPR19274] to DRP1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ICC/IF, IP, Flow Cyt, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Mouse DRP1 aa 1-350. The exact sequence is proprietary.
    Database link: Q8K1M6

  • Positive control
    • WB: Human fetal kidney, rat brain, rat heart and mouse brain lysates; A549, U-2 OS, HeLa, Jurkat, HEK-293, HCT 116, PC-12 and NIH/3T3 whole cell lysates. IHC-P: Mouse cerebrum and rat cerebellum tissues. ICC/IF: HeLa and NIH/3T3 cells. Flow Cyt: NIH/3T3 cells. IP: HeLa whole cell lysate.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab184247 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000. Detects a band of approximately 83 kDa (predicted molecular weight: 83 kDa).
ICC/IF 1/250.
IP 1/30.
Flow Cyt 1/70.
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

IHC is recommended for rat and mouse only.

Target

  • Function
    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.
  • Tissue specificity
    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.
  • Involvement in disease
    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.
  • Sequence similarities
    Belongs to the dynamin family.
    Contains 1 GED domain.
  • Domain
    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.
  • Post-translational
    modifications
    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.
  • Cellular localization
    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.
  • Information by UniProt
  • Database links
  • Alternative names
    • DLP1 antibody
    • dnm1l antibody
    • DNM1L_HUMAN antibody
    • Dnm1p/Vps1p-like protein antibody
    • dnml1 antibody
    • DRP1 antibody
    • DVLP antibody
    • Dymple antibody
    • Dynamin 1 like antibody
    • Dynamin family member proline-rich carboxyl-terminal domain less antibody
    • Dynamin like protein antibody
    • Dynamin related protein 1 antibody
    • Dynamin-1-like protein antibody
    • Dynamin-like protein 4 antibody
    • Dynamin-like protein antibody
    • Dynamin-like protein IV antibody
    • Dynamin-related protein 1 antibody
    • DYNIV 11 antibody
    • EMPF antibody
    • EMPF1 antibody
    • FLJ41912 antibody
    • HdynIV antibody
    • VPS1 antibody
    see all

Images

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling DRP1 with ab184247 at 1/250 dilution, followed by Goat anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasm staining on HeLa cell line. The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [EPR19274] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab184247 at 1/250 dilution followed by ab150120  at 1/1000 dilution.
    -ve control 2: ab7291  at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling DRP1 with ab184247 at 1/70 dilution (red) compared with a Rabbit IgG,monoclonal -Isotype Control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.

  • All lanes : Anti-DRP1 antibody [EPR19274] (ab184247) at 1/1000 dilution

    Lane 1 : A549 (Human lung carcinoma cell line) whole cell lysate
    Lane 2 : U-2 OS (Human bone osteosarcoma epithelial cell line) whole cell lysate
    Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
    Lane 5 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
    Lane 6 : HCT 116 (Human colorectal carcinoma cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 83 kDa
    Observed band size: 83 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1 and 2: 3 minutes; Lane 3: 30 seconds; Lane 4,5 and 6: 8 seconds.

    DRP1 can be SUMOylated, as described in the literature (PMID: 19638400).

  • Anti-DRP1 antibody [EPR19274] (ab184247) at 1/1000 dilution + Human fetal kidney lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/100000 dilution

    Predicted band size: 83 kDa
    Observed band size: 83 kDa


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-DRP1 antibody [EPR19274] (ab184247) at 1/1000 dilution

    Lane 1 : Rat brain lysate
    Lane 2 : Rat heart lysate
    Lane 3 : Mouse brain lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 83 kDa
    Observed band size: 83 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Lane 1: 2 seconds; Lane 2: 8 seconds; Lane 3: 3 seconds.

  • All lanes : Anti-DRP1 antibody [EPR19274] (ab184247) at 1/1000 dilution

    Lane 1 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 83 kDa
    Observed band size: 83 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling DRP1 with ab184247 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on mouse cerebrum is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded Rat cerebellum tissue labeling DRP1 with ab184247 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on rat cerebellum is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling DRP1 with ab184247 at 1/250 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasm staining on NIH/3T3 cell line. The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [EPR19274] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab184247 at 1/250 dilution followed by ab150120  at 1/1000 dilution.
    -ve control 2: ab7291 at 1/1000 dilution followed by ab150077  at 1/1000 dilution.

  • DRP1 was immunoprecipitated from 1mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab184247 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab184247 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/10000 dilution.
    Lane 1: HeLa whole cell lysate 10µg (Input).
    Lane 2: ab184247 IP in HeLa whole cell lysate.
    Lane 3: Rabbit IgG,monoclonal [EPR19274]-Isotype Control  (ab172730) instead of ab184247 in HeLa whole cell lysate.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 5 seconds.

    Note: DRP1 can be SUMOylated, as described in the literature (PMID: 19638400).

References

This product has been referenced in:
  • Yan X  et al. Lup-20(29)-en-3ß,28-di-yl-nitrooxy acetate affects MCF-7 proliferation through the crosstalk between apoptosis and autophagy in mitochondria. Cell Death Dis 9:241 (2018). Read more (PubMed: 29445224) »
  • Nguyen HT  et al. Small-Vessel Vasculopathy Due to Aberrant Autophagy in LAMP-2 Deficiency. Sci Rep 8:3326 (2018). Read more (PubMed: 29463847) »

See all 5 Publications for this product

Customer reviews and Q&As

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Application
Western blot
Sample
Human Cell lysate - whole cell (Leukemic Cancer Cell lines)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
20 µg
Specification
Leukemic Cancer Cell lines
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C
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Submitted Dec 13 2017

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