Overview

  • Product nameAnti-Doublecortin antibody
    See all Doublecortin primary antibodies
  • Description
    Rabbit polyclonal to Doublecortin
  • SpecificityPlease note: Low dilutions of this antibody can cause high background. Please use as high a dilution as possible. Optimal working dilutions are batch dependent.
  • Tested applicationsSuitable for: WB, ICC, ICC/IF, IHC-FrFl, IHC-FoFr, IHC-Fr, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Cat, Human, Cynomolgus Monkey, Quail, Rhesus monkey
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 300 to the C-terminus of Human Doublecortin.

    (Peptide available as ab19804.)

  • Positive control
    • This antibody gave a positive signal in Brain (Mouse) Tissue Lysate - normal tissue, 0 days old

Properties

Applications

Our Abpromise guarantee covers the use of ab18723 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 Use a concentration of 1 µg/ml. Detects a band of approximately 45 kDa (predicted molecular weight: 40-45 kDa).
ICC Use at an assay dependent concentration.
ICC/IF Use a concentration of 1 - 5 µg/ml.
IHC-FrFl 1/1000. Please note: Low dilutions of this antibody can cause high background. Please use as high a dilution as possible. Optimal working dilutions are batch dependent.
IHC-FoFr 1/1000.
IHC-Fr 1/2000 - 1/7000.
IHC-P Use a concentration of 0.1 - 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

  • FunctionSeems 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 specificityHighly 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 diseaseDefects 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 similaritiesContains 2 doublecortin domains.
  • Cellular localizationCytoplasm.
  • 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

Anti-Doublecortin antibody images

  • ab18723 staining Doublecortin in mouse embryonic 15 day brain tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with paraformaldehyde, permeablized with 0.5% Triton X-100 in PBS, blocked with 10% serum for 1 hour at 25°C and antigen retrieval was by heat mediation in citrate buffer, pH 6. The sample was incubated with primary antibody (1/500 in PBS + 0.1% Triton X-100 + 1% serum) for 16 hours at 25°C. An Alexa Fluor® 594-conjugated donkey anti-rabbit IgG polyclonal (1/700) was used as the secondary antibody.

    See Abreview

  • IHC-Fr image of Doublecortin staining in Mouse adult dentate gyrus sections using ab18723(1:100). The sections were fixed in paraformaldehyde and permeabilized using 1x TBST. The sections were then blocked using 10% donkey serum for 1 hour at 25°C. ab18723 was diluted 1:100 and incubated with the sections for 12 hours at 25°C. The secondary antibody used was Donkey anti-rabbit conjugated to Cy3 Dye (1:500). DAPI was used to stain the nuclei.

    See Abreview

  • ab18723 staining 6 week rat brain tissue dentate gyrus (DG) by IHC-P using rabbit-specific EXPOSE IHC detection kit (ab80437). Formalin fixed paraffin embedded tissue sections were pre-treated using heat mediated antigen retrieval (using a pressure cooker) with sodium citrate buffer (pH6) for 30 mins. The section was incubated with ab18723, 0.1µg/ml, for 1 hour at room temperature. DAB was used as the chromogen and the section was counterstained with haematoxylin and mounted with DPX.

  • ab18723 at 1/200 staining mouse E18 body T/S spinal cord tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retireval step was perfomed before incubation with the antibody for 24 hours. A biotinylated goat antibody was used as the secondary.

    See Abreview

  • Doublecortin antibody (ab18723; green) labeling cell extensions consistent with dendrite morphology in 4 day old cultures. Preincubation of ab18723 with its immunising peptide (ab19804) quenched immunostaining (see review).

    Dorsal root ganglion explants were dissected from 16 day-old rat embryos and cultured for 4 days in vitro with Neurobasal Medium containing 10% fetal calf serum and B27 supplement. Immunocytochemistry: All steps were performed in PBS. Cells or explants were fixed in 4% PFA for 15min, permeabilised with 0.1% TX100 for 10min and blocked with 5% BSA, 0.1% TX100 for 45min. ab18723 was incubated at 5µg/ml for 12h in 5% BSA, 0.1% TX100 at 4°C. For peptide blocking experiments preincubation of the peptide (ab19804; 250µg/ml) and antibody (5µg/ml ) was performed for 2h at 37°C. Cultures were washed (3x) of primary antibody solution. Goat anti-rabbit AlexaFluor 488 was used as secondary antibody (1/400) in 5% BSA, 0.1% TX100 for 2h at R

  • Doublecortin expression in the dentate gyrus of a 1 month-old mouse brain. Doublecortin staining using ab18723 (1/500) in the dentate gyrus of a 1 month-old mouse brain. The mouse has been perfused with paraformaldehyde 4% (50ml). After dissection, the brain has been incubated overnight in sucrose 20%, embedded in OCT and cryosectioned (10 µm). No antigen retrieval was used. The secondary antibody used was a non-Abcam Goat anti-rabbit Alexa488.
  • IHC image of ab18723 staining in rat 6 week brain formalin fixed paraffin embedded tissue section, performed on a Leica BondTM 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 ab18723, 0.1µg/ml, 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.

    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.
  • Immunohistochemistical staining (on formaldehyde/PFA-fixed paraffin-embedded sections) of Doublecortin antibody - Neuronal Marker (ab18723) on Quail Tissue sections (E6/7 brain (Saggital section). Antigen retrieval step: Heat mediated. Blocking step: 1% BSA for 10 mins at RT. Primary Antibody ab18723 incubated at 1/400 for 2 hours RT. Secondary Antibody: Biotin conjugated goat anti rabbit Ig (1/300).
  • ab18723 at 1/2000 staining mouse brain svr: progenitor olfactory neurones by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step was performed before incubation with the antibody for 16 hours. An Alexa-Fluor ® 488 conjugated goat antibody was used as the secondary (green). The tissue was also stained for Ki67 (shown in red).

    The MIP image was derived from Apotome-generated Z-stacks from the greyscale image of each of the channels in the MIP.

    See Abreview

  • ab18723 staining Doublecortin in mouse dentate gyrus hippocampal tissue section by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were incubated with primary antibody 1/1000 for 16 hours at 4°C.  An Alexa Fluor®488 conjugated goat polyclonal to rabbit IgG at 1/400 dilution was used as secondary.

    See Abreview

  • Anti-Doublecortin antibody (ab18723) at 1 µg/ml + Mouse brain tissue lysate - total protein (0 days) (ab7188) at 10 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 40-45 kDa
    Observed band size : 45 kDa (why is the actual band size different from the predicted?)

References for Anti-Doublecortin antibody (ab18723)

This product has been referenced in:
  • Fu S  et al. Aberrant adult Neurogenesis in the subventricular zone-rostral migratory stream-olfactory bulb system following subchronic manganese exposure. Toxicol Sci N/A:N/A (2016). Read more (PubMed: 26794142) »
  • Siopi E  et al. Anxiety- and Depression-Like States Lead to Pronounced Olfactory Deficits and Impaired Adult Neurogenesis in Mice. J Neurosci 36:518-31 (2016). Read more (PubMed: 26758842) »

See all 82 Publications for this product

Product Wall

Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Mouse Tissue sections (BRAIN)
Antigen retrieval step None
Permeabilization Yes - 4% paraform
Specification BRAIN
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Fixative Paraformaldehyde
Username

Ms. kyungjoo seong

Verified customer

Submitted Nov 30 2016

Application Immunocytochemistry
Sample Mouse Cell (Neuron)
Permeabilization No
Specification Neuron
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Fixative Paraformaldehyde
Username

Ms. kyungjoo seong

Verified customer

Submitted Nov 01 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (Brain)
Permeabilization Yes - 0.3% triton x-100
Specification Brain
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jun 01 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Mouse Tissue sections (brain)
Antigen retrieval step None
Permeabilization Yes - triton 0.5%
Specification brain
Blocking step mix bsa (1%) and dk serum (5%) as blocking agent for 2 hour(s) and 0 minute(s) · Temperature: 22°C
Fixative Paraformaldehyde
Username

Ms. Francoise Geffroy

Verified customer

Submitted Apr 21 2016

Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Cell lysate - whole cell (liver)
Specification liver
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted Dec 31 2014

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (10% SDS PAGE)
Sample Mouse Tissue lysate - whole (Mouse brain)
Specification Mouse brain
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jun 06 2014

Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate buffer, pH 6
Sample Mouse Tissue sections (Embryonic 15 day brain)
Specification Embryonic 15 day brain
Permeabilization Yes - 0.5% Triton X-100 in PBS
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted May 16 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 18°C
Sample Human Cell (hippocampal progenitor cells)
Specification hippocampal progenitor cells
Permeabilization Yes - 0.3% Triton X-100
Fixative Paraformaldehyde
Username

Dr. Aleksandra Maruszak

Verified customer

Submitted Mar 31 2014

ab77450 is raised using the same immunogen as ab18723 and therefore the products are essentially identical. The only difference between these products is that batches sold under ab77450 have passed QC testing in Western Blot and ICC/IF however, in our ...

Read More
Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 40 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 22°C
Sample Mouse Cell (neuroblast)
Specification neuroblast
Permeabilization Yes - 0.2% TritonX 100
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted Oct 21 2013

1-10 of 44 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"