Overview

  • Product name
    Anti-Doublecortin antibody [EPR19997]
    See all Doublecortin primary antibodies
  • Description
    Rabbit monoclonal [EPR19997] to Doublecortin
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, WB, IHC-Fr, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human Doublecortin aa 250 to the C-terminus. The exact sequence is proprietary.
    Database link: O43602-2

  • Positive control
    • WB: Human fetal brain and brain lysates; Mouse and rat brain lysates; SH-SY5Y whole cell lysate. IHC-P: Mouse and rat adult hippocampus tissues. IHC-Fr: Rat adult hippocampus tissue. ICC/IF: SH-SY5Y cells.
  • 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 ab207175 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
IHC-P 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa).
IHC-Fr 1/100.

Antigen retrieval: Heated citrate solution (10mM citrate PH 6.0 + 0.05% Tween-20).

ICC/IF 1/250.

Target

  • Function
    Seems to be required for initial steps of neuronal dispersion and cortex lamination during cerebral cortex development. May act by competing with the putative neuronal protein kinase DCAMKL1 in binding to a target protein. May in that way participate in a signaling pathway that is crucial for neuronal interaction before and during migration, possibly as part of a calcium ion-dependent signal transduction pathway. May be part with LIS-1 of an overlapping, but distinct, signaling pathways that promote neuronal migration.
  • Tissue specificity
    Highly expressed in neuronal cells of fetal brain (in the majority of cells of the cortical plate, intermediate zone and ventricular zone), but not expressed in other fetal tissues. In the adult, highly expressed in the brain frontal lobe, but very low expression in other regions of brain, and not detected in heart, placenta, lung, liver, skeletal muscles, kidney and pancreas.
  • Involvement in disease
    Defects in DCX are the cause of lissencephaly X-linked type 1 (LISX1) [MIM:300067]; also called X-LIS or LIS. LISX1 is a classic lissencephaly characterized by mental retardation and seizures that are more severe in male patients. Affected boys show an abnormally thick cortex with absent or severely reduced gyri. Clinical manifestations include feeding problems, abnormal muscular tone, seizures and severe to profound psychomotor retardation. Female patients display a less severe phenotype referred to as 'doublecortex'.
    Defects in DCX are the cause of subcortical band heterotopia X-linked (SBHX) [MIM:300067]; also known as double cortex or subcortical laminar heterotopia (SCLH). SBHX is a mild brain malformation of the lissencephaly spectrum. It is characterized by bilateral and symmetric plates or bands of gray matter found in the central white matter between the cortex and cerebral ventricles, cerebral convolutions usually appearing normal.
    Note=A chromosomal aberration involving DCX is found in lissencephaly. Translocation t(X;2)(q22.3;p25.1).
  • Sequence similarities
    Contains 2 doublecortin domains.
  • Cellular localization
    Cytoplasm.
  • Information by UniProt
  • Database links
  • Alternative names
    • DBCN antibody
    • Dbct antibody
    • DC antibody
    • DCX antibody
    • DCX_HUMAN antibody
    • Doublecortex antibody
    • Doublin antibody
    • FLJ51296 antibody
    • Lis X antibody
    • Lis-X antibody
    • Lissencephalin X antibody
    • Lissencephalin-X antibody
    • Lissencephaly X linked antibody
    • Lissencephaly X linked doublecortin antibody
    • LISX antibody
    • Neuronal migration protein doublecortin antibody
    • OTTHUMP00000023859 antibody
    • OTTHUMP00000023860 antibody
    • OTTHUMP00000216315 antibody
    • OTTHUMP00000216316 antibody
    • SCLH antibody
    • XLIS antibody
    see all

Images

  • IHC image of Doublecortin staining in a formalin fixed, paraffin embedded normal rat hippocampus (SVZ region) tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab207175 at 1/500 dilution for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. As a negative control (inset), an identical assay was performed without adding the primary antibody.

     

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Immunohistochemical analysis of paraffin-embedded mouse adult hippocampus tissue labeling Doublecortin with ab207175 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on mouse hippocampal dentate gyrus is observed [PMID:23690918] [PMID:16814555]. 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 adult hippocampus tissue labeling Doublecortin with ab207175 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on rat hippocampal dentate gyrus is observed [PMID:23690918] [PMID:16814555]. 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 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen rat adult hippocampus tissue labeling Doublecortin with ab207175 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Cytoplasmic staining on rat hippocampal dentate gyrus is observed [PMID:23690918] [PMID:16814555].  The nuclear counterstain is DAPI (blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Immunofluorescent analysis of 100% Methanol-fixed SH-SY5Y (Human neuroblastoma cell line from bone marrow) cells labeling Doublecortin with ab207175 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on SH-SY5Y cell line.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • All lanes : Anti-Doublecortin antibody [EPR19997] (ab207175) at 1/10000 dilution

    Lane 1 : Human fetal brain lysate
    Lane 2 : Human brain lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 40 kDa
    Observed band size: 40 kDa


    Exposure time: 3 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The higher molecular weight band is phosphorylated form and the lower molecular weight band is non-phosphorylated form. (PMID: 17178868).

    Doublecortin is highly expressed during embryonic development and downregulated in adult tissues. (PMID: 10399933).

  • All lanes : Anti-Doublecortin antibody [EPR19997] (ab207175) at 1/10000 dilution

    Lane 1 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate
    Lane 2 : Mouse brain lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 40 kDa
    Observed band size: 40 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure times: Lane 1: 15 seconds; Lane 2: 3 minutes.

    The higher molecular weight band is phosphorylated form and the lower molecular weight band is non-phosphorylated form. (PMID: 17178868).

    Doublecortin is highly expressed during embryonic development and downregulated in adult tissues. (PMID: 10399933).

  • All lanes : Anti-Doublecortin antibody [EPR19997] (ab207175) at 1/1000 dilution

    Lane 1 : Human fetal brain lysate
    Lane 2 : Human fetal heart lysate
    Lane 3 : Mouse brain lysate
    Lane 4 : Mouse heart lysate
    Lane 5 : Rat brain lysate
    Lane 6 : Rat heart 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: 40 kDa
    Observed band size: 40 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure times: Lane 1-4: 3 minutes; Lane 5/6: 10 seconds.

    The observed expression profile is consistent with what has been described in the literature (PMID: 10550327).

References

This product has been referenced in:
  • Yu L  et al. BMP9 stimulates joint regeneration at digit amputation wounds in mice. Nat Commun 10:424 (2019). Read more (PubMed: 30723209) »
  • Corbetta C  et al. Altered function of the glutamate-aspartate transporter GLAST, a potential therapeutic target in glioblastoma. Int J Cancer N/A:N/A (2018). Read more (PubMed: 30418668) »
See all 2 Publications for this product

Customer reviews and Q&As

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1-2 of 2 Abreviews

Application
Immunohistochemistry free floating
Sample
Zonotrichia leucophrys Tissue sections (Brain)
Specification
Brain

Abcam user community

Verified customer

Submitted Apr 30 2019

Application
Immunocytochemistry/ Immunofluorescence
Sample
Cynomolgus monkey Cell (Embryonic cell of cortex)
Permeabilization
Yes - Triton 0,5%
Specification
Embryonic cell of cortex
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Feb 28 2019

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