Overview

  • Product nameAnti-Nestin antibody [401]
    See all Nestin primary antibodies
  • Description
    Mouse monoclonal [401] to Nestin
  • Tested applicationsICC/IF, ICC, IHC-Fr, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat

    Does not react with

    Human
  • Immunogen

    Nestin purified from embryonic rat spinal cord.

  • Positive control
    • MEF1 whole cell lysate

Properties

Applications

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.

Application Abreviews Notes
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.

Target

  • FunctionRequired 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.
  • Tissue specificityCNS stem cells.
  • Sequence similaritiesBelongs to the intermediate filament family.
  • Developmental stageUpon terminal neural differentiation, nestin is down-regulated and replaced by neurofilaments.
  • Post-translational
    modifications
    Constitutively phosphorylated. This increases during mitosis when the cytoplasmic intermediate filament network is reorganized.
  • Information by UniProt
  • Database links
  • Alternative names
    • ESTM 46 antibody
    • ESTM 46 antibody
    • FLJ 21841 antibody
    • FLJ 21841 antibody
    • FLJ21841 antibody
    • Intermediate filament protein antibody
    • Intermediate filament protein antibody
    • Nbla00170 antibody
    • Nes antibody
    • Nes antibody
    • NEST_HUMAN antibody
    • Nestin antibody
    see all

Anti-Nestin antibody [401] images

  • Anti-Nestin antibody [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

  • 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
  • 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.

References for Anti-Nestin antibody [401] (ab11306)

This product has been referenced in:
  • Liu H  et al. p53 Regulates Neural Stem Cell Proliferation and Differentiation via BMP-Smad1 Signaling and Id1. Stem Cells Dev : (2013). Read more (PubMed: 23199293) »
  • Daniel C  et al. Nestin expression in repopulating mesangial cells promotes their proliferation. Lab Invest 88:387-97 (2008). Read more (PubMed: 18268475) »

See all 13 Publications for this product

Product Wall

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

Submitted Jun 18 2012

Thank you for contacting us. Unfortunately the exact immunogen sequence for this product is proprietary. However, I may tell that it was derived from within residues 100 - 200 of Mouse Pericentrin. I hope this information is helpful to you. Ple...

Read More
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

Submitted 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

Submitted Feb 02 2010

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