Overview

  • Product nameAnti-Nestin antibody [2Q178]
    See all Nestin primary antibodies
  • Description
    Mouse monoclonal [2Q178] to Nestin
  • Tested applicationsICC/IF, Electron Microscopy, Flow Cyt, IHC-FoFr, IHC-P, IHC-Fr, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat

    Does not react with

    Sheep, Cat, Human, Monkey
  • Immunogen

    Full length native Nestin protein purified from embryonic rat spinal cord.

  • Positive control
    • Mouse or Rat embryonic brain.

Properties

Applications

Our Abpromise guarantee covers the use of ab6142 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 1/20 - 1/200. Cells fixed with 4% paraformaldehyde buffered with 50mM sodium borate at pH 9.5.
Electron Microscopy Use a concentration of 0.1 - 1 µg/ml. Tissue fixed with 4% paraformaldehyde at pH 10.0 or 4% paraformaldehyde with 0.1% glutaraldehyde at pH 7.4.
Flow Cyt Use at an assay dependent dilution. PubMed: 21092738
IHC-FoFr Use at an assay dependent dilution. PubMed: 18562299
IHC-P Use a concentration of 0.1 - 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. Tissue fixed with 4% paraformaldehyde at pH 7.4 for light microscopy.
IHC-Fr Use a concentration of 0.1 - 1 µg/ml. Tissue fixed with 4% paraformaldehyde at pH 7.4 for light microscopy.
WB Use at an assay dependent dilution. Predicted molecular weight: 200 kDa. Block with milk or BSA but do not dilute primary antibody in buffer containing milk.

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
    • FLJ 21841 antibody
    • FLJ21841 antibody
    • Intermediate filament protein antibody
    • Nbla00170 antibody
    • nes antibody
    • NEST_HUMAN antibody
    • Nestin antibody
    see all

Anti-Nestin antibody [2Q178] images

  • ab6142 at 1/500 dilution staining Nestin in rat brain tissue (green) by immunohistochemistry (frozen sections). Sections were methanol fixed, permeabilized in 0.1% Triton X-100 prior to blocking in 2.5% BSA for 16 hours at 4°C and then incubated with ab6142, for 1 hour at 37°C. Alexa fluor® 680 goat polyclonal to mouse Ig, diluted 1/1000 was used as the secondary antibody.

    See Abreview

  • ab6142 staining adult mouse brain tissue section by Immunohistochemistry (Formalin/PFA-fixed, paraffin embedded sections). Tissue underwent fixation in paraformaldehyde, heat mediated antigen retrieval in Sodium Citrate, permeabilization in 1% Triton buffer and blocking in 10% serum for 1 hour at 25°C. The primary antibody, diluted 1/200 (PBS, 2% Donkey serum, 0.2% Triton) for 16 hours at 4°C.  An Alexa Fluor® 488 conjugated donkey polyclonal to mouse Ig, diluted 1/500 was used as the secondary.

    See Abreview

  • ab6142 at 1/200 dilution staining Nestin in mouse Cor1 neural stem cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed using 4%PFA for 15mins at RT. After x3 PBS rinses over 15mins, cells were blocked/permeabilised using primary Antibody dilution buffer for 15mins befpre application of primary Ab diluted in same buffer. Secondary conjugate was diluted in same buffer, with addition of 1/10K Hoechst, to stain nuclei. After 60mins cells were rinsed x3 over 15mins before coverslipping using Mowiol containing DABCO and azide.

    See Abreview

  • ab6142 staining mouse embryonic stem cells by immunocytochemistry/ immunofluorescence. Cells were PFA fixed and permeabilized in 0.4% Triton X-100 prior to blocking in 5% serum for 1 hour at 25°C. The primary antibody was diluted 1/500 and incubated with the sample for 20 hours at 4°C. Alexa fluor® 568 goat polyclonal to mouse Ig, diluted 1/400, was used as the secondary antibody.

    See Abreview

References for Anti-Nestin antibody [2Q178] (ab6142)

This product has been referenced in:
  • Schmidt-Edelkraut U  et al. Zac1 regulates cell cycle arrest in neuronal progenitors via Tcf4. Mol Cell Biol 34:1020-30 (2014). Read more (PubMed: 24396065) »
  • Mastronuzzi A  et al. Large cell anaplastic medulloblastoma metastatic to the scalp: tumor and derived stem-like cells features. BMC Cancer 14:262 (2014). IF ; Human . Read more (PubMed: 24739212) »

See all 26 Publications for this product

Product Wall

Thank you for contacting us.
I can confirm the isotype controls suggested are suitable for the antibodies requested.
The difference in price between ab91353 and ab81197 lies in the volume difference (the volume of ab91353 doubles ab81197’s vo...

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Application Western blot
Sample Apteronotus leptorhynchus Tissue lysate - whole (Brain)
Loading amount 50 µg
Specification Brain
Gel Running Conditions Reduced Denaturing (4-15%)
Blocking step Milk as blocking agent for 14 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Nov 27 2012

Vielen Dank für Ihre Anfrage.

Ich stimme Ihnen zu, dass man Mausantikörper auf Mausgeweben generell nicht empfiehlt, da der sekundaere Antikörper auch die endogenes Ig erkennt und somit ein starker Hintergrund auftritt.
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Gracias por contactar con nosotros.

Lamento mucho las molestias ocasionadas.

Sin embargo te rogaría que por favor me confirmes si habéis centrifugado el vial antes de abrirlo, y comprobado que el líquido no se q...

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Thank you very much for your interest in anti-Nestin antibodies.

To our knowledge our anti-Nestin products have not been tested in bovine. Therefore, I can offer a discount off a future purchase if you buy an anti-Nestin antibody now, test it...

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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 (Brain)
Specification Brain
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate buffer
Permeabilization No
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
Username

Dr. R Berahovich

Verified customer

Submitted Dec 01 2011

Thank you for your call today, and I apologize for the trouble with these codes! I have created new codes which will expire on March 1, 2012. I will go through your Abreviews and put these new codes in our internal notes so that the discounts can be ap...

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Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Apteronotus leptorhynchus Tissue sections (Spinal cord, brain)
Specification Spinal cord, brain
Fixative Paraformaldehyde
Permeabilization Yes - 0.3% Triton X-100
Blocking step 3% sheep serum, 1% BSA, 1% teleostean gelatine in PBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C
Username

Dr. Ruxandra Sirbulescu

Verified customer

Submitted Nov 14 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Flow Cytometry
Sample Mouse Cell (Spleen)
Specification Spleen
Preparation Cell harvesting/tissue preparation method: 4% PFA was used to fixed the cells for 30 mins at room temperature.
Sample buffer: 1% BSA PBS was used as wash buffer
Fixation Paraformaldehyde
Permeabilization Yes - 0.5% Triton X-100
Username

Abcam user community

Verified customer

Submitted Jul 20 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (mouse ES cells)
Specification mouse ES cells
Fixative Paraformaldehyde
Permeabilization Yes - 0.4% Triton X-100
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Oct 20 2009

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