概述

  • 产品名称
    Anti-Nestin抗体[大鼠-401]
    参阅全部 Nestin 一抗
  • 描述
    小鼠单克隆抗体[大鼠-401] to Nestin
  • 经测试应用
    适用于: ICC/IF, ICC, IHC-Fr, WB, IHC-Pmore details
  • 种属反应性
    与反应: Mouse, Rat
    不与反应: Human
  • 免疫原

    Nestin purified from embryonic rat spinal cord.

  • 阳性对照
    • WB: MEF1 whole cell lysate. ICC/IF: PC12 cells. IHC-P: 6 week old rat brain FFPE tissue sections.
  • 常规说明

    This antibody clone is manufactured by Abcam.

    The Protocols tab contains a Mouse on Mouse staining protocol with recommendations when using a mouse monoclonal antibody to stain mouse tissues and tips for reducing background.

性能

应用

Our Abpromise guarantee covers the use of ab11306 in the following tested applications.

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

应用 Ab评论 说明
ICC/IF Use a concentration of 10 µg/ml.
ICC Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 170, 350 kDa (predicted molecular weight: 177 kDa). The use of BSA as blocking agent is not recommended.
IHC-P Use a concentration of 1 - 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

靶标

  • 功能
    Required for brain and eye development. Promotes the disassembly of phosphorylated vimentin intermediate filaments (IF) during mitosis and may play a role in the trafficking and distribution of IF proteins and other cellular factors to daughter cells during progenitor cell division. Required for survival, renewal and mitogen-stimulated proliferation of neural progenitor cells.
  • 组织特异性
    CNS stem cells.
  • 序列相似性
    Belongs to the intermediate filament family.
  • 发展阶段
    Upon terminal neural differentiation, nestin is down-regulated and replaced by neurofilaments.
  • 翻译后修饰
    Constitutively phosphorylated. This increases during mitosis when the cytoplasmic intermediate filament network is reorganized.
  • Information by UniProt
  • 数据库链接
  • 别名
    • ESTM 46 antibody
    • FLJ 21841 antibody
    • FLJ21841 antibody
    • Intermediate filament protein antibody
    • Nbla00170 antibody
    • nes antibody
    • NEST_HUMAN antibody
    • Nestin antibody
    see all

Anti-Nestin antibody [Rat-401] 图像

  • Immunocytochemical analysis of Rat neural stem cells from the hippocampus, labelling Nestin with ab11306. Samples were fixed in paraformaldehyde, permeablized in triton x-100 and blocked in 1% BSA for 40 minutes at room temperature. Incubation with ab11306 diluted 1/133 for 12 hours at 4°C. Secondary antibody was a polyclonal goat anti mouse FITC diluted 1/200.

    See Abreview

  • Anti-Nestin antibody [Rat-401] (ab11306) at 1 µg/ml + MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) (ab65485) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 177 kDa


    Exposure time : 20 minutes
  • ab11306 staining Nestin in rat lateral ventricle (subventricular zone)  sections by IHC-P (formaldehyde-fixed paraffin-embedded sections). Tissue samples were fixed with formaldehyde and blocked with  1% BSA for 10 minutes at room temperature. Antigen retrieval was by heat mediation in citric acid (pH6). Samples were incubated with primary antibody 1/200 (TBS/BSA/azide pH7.5) for 24 hours. A Biotin-conjugated Goat polycolonal to mouse IgG, dilution 1/200, was used as secondary antibody. The image shows clear staining in cells of the rat subventricular zone of the lateral ventricle.

    See Abreview

  • ICC/IF image of ab11306 stained PC12 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab11306, 10µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) PC12 cells at 10µg/ml.
  • IHC image of Nestin staining in 6 week old rat 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 ab11306, 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

Anti-Nestin antibody [Rat-401] (ab11306)参考文献

This product has been referenced in:
  • Fang L  et al. Jumonji AT-rich interactive domain 1B overexpression is associated with the development and progression of glioma. Int J Mol Med 38:172-82 (2016). WB ; Human . Read more (PubMed: 27246838) »
  • Lai Q  et al. NMDA receptors promote neurogenesis in the neonatal rat subventricular zone following hypoxic-ischemic injury. Mol Med Rep 13:206-12 (2016). Rat . Read more (PubMed: 26548659) »

See all 27 Publications for this product

Product Wall

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium Citrate
Permeabilization
Yes - Brij
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 37°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Aug 21 2015

Application
Immunohistochemistry (Frozen sections)
Blocking step
Serum as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Sample
Rat Tissue sections (Brain)
Specification
Brain
Permeabilization
No
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Apr 10 2015

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Rat Tissue sections (Brain)
Specification
Brain
Fixative
Paraformaldehyde
Antigen retrieval step
None
Permeabilization
No
Username

Dr. Sophie Pezet

Verified customer

提交于 Jun 18 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry
Sample
Rat Cell (neural stem cell,hippocampus)
Specification
neural stem cell,hippocampus
Fixative
Paraformaldehyde
Permeabilization
Yes - triton x-100
Blocking step
BSA as blocking agent for 40 minute(s) · Concentration: 1% · Temperature: RT°C
Username

Dr. san san

Verified customer

提交于 Aug 18 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (adult rat svz of lateral ventricle)
Specification
adult rat svz of lateral ventricle
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid pH6. Not a buffer
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: rt°C
Username

Mr. Carl Hobbs

Verified customer

提交于 Feb 02 2010

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

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