製品の概要

  • 製品名
    Anti-Doublecortin antibody
    Doublecortin 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to Doublecortin
  • 特異性
    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.
  • アプリケーション
    適用あり: WB, ICC, ICC/IF, IHC-FrFl, IHC-FoFr, IHC-Fr, IHC-Pmore details
  • 種交差性
    交差種: Mouse, Rat, Chicken, Cat, Human, Cynomolgus monkey, Quail, Rhesus monkey
  • 免疫原

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

    (Peptide available as ab19804.)

  • ポジティブ・コントロール
    • This antibody gave a positive signal in Brain (Mouse) Tissue Lysate - normal tissue, 0 days old ICC-IF: SHSY5Y cells.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

ターゲット情報

  • 機能
    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.
  • 組織特異性
    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.
  • 関連疾患
    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).
  • 配列類似性
    Contains 2 doublecortin domains.
  • 細胞内局在
    Cytoplasm.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 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

画像

  • 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.

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  • 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.

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  • 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.

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  • ab18723 stained in SHSY5Y cells. Cells were fixed with 100% methanol (5 min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab18723 at 5 µg/ml and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150120 (pseudo-colored red) and ab150081 (colored green) used at 1 ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  • 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) 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.

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  • 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?)

参考文献

This product has been referenced in:
  • Zhu G  et al. Endoplasmic reticulum stress mediates distinct impacts of sevoflurane on different subfields of immature hippocampus. J Neurochem 142:272-285 (2017). Read more (PubMed: 28444766) »
  • Song S  et al. Effects of an Inhibitor of Monocyte Recruitment on Recovery from Traumatic Brain Injury in Mice Treated with Granulocyte Colony-Stimulating Factor. Int J Mol Sci 18:N/A (2017). IHC-Fr ; Mouse . Read more (PubMed: 28671601) »

See all 125 Publications for this product

レビューと Q&A

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Bird Tissue sections (Serinus canaria forma domestica brain)
Antigen retrieval step
None
Permeabilization
Yes - triton-x 0,1% in Tris buffered saline
Specification
Serinus canaria forma domestica brain
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 24°C
Fixative
Paraformaldehyde
Username

Laura Vandries

Verified customer

投稿 Jul 13 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Sodium Citrate, 0.05% Tween 20, pH 6.0
Permeabilization
No
Specification
brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Dec 16 2016

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

Kyungjoo Seong

Verified customer

投稿 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

Kyungjoo Seong

Verified customer

投稿 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

投稿 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

Mrs. Francoise Geffroy

Verified customer

投稿 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

投稿 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

投稿 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

投稿 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

投稿 Mar 31 2014

1-10 of 46 Abreviews or Q&A

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

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