Overview

  • Product name
    Anti-Nestin antibody [10C2]
    See all Nestin primary antibodies
  • Description
    Mouse monoclonal [10C2] to Nestin
  • Specificity
    Reacts exclusively with human nestin protein.
  • Tested applications
    Suitable for: ICC/IF, IHC-Fr, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    A recombinant 150 amino acid fragment from human Nestin conjugated to glutathione S transferase.

  • General notes

    Monoclonal antibody 10C2 was derived by fusion of P3x63Ag8.653 myeloma cells with spleen cells from a BALB/c mouse immunized with a 150-amino-acid fragment from the cloned human nestin, conjugated to glutathione S-transferase.

Properties

Applications

Our Abpromise guarantee covers the use of ab22035 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 1/100 - 1/200. PubMed: 19147502

1/1000.

IHC-Fr 1/100 - 1/200.
IHC-P Use at an assay dependent concentration.
WB 1/1000 - 1/5000. Predicted molecular weight: 176 kDa.

1/1000 - 1/5000 using chemiluminescent detection. Predicted molecular weight: 176 kDa.

Target

  • Function
    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.
  • Tissue specificity
    CNS stem cells.
  • Sequence similarities
    Belongs to the intermediate filament family.
  • Developmental stage
    Upon 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

Images

  • ab22035 staining human brain tissue cells by ICC/IF.  Cells were PFA fixed and permeabilized in Triton X-100 prior to blocking in 1% BSA for 30 minutes at 21°C.  The primary antibody was diluted 1/100 and incubated with the samples for 1 hour at 21°C.  A FITC conjugated goat anti-mouse IgG antibody, diluted 1/250, was used as the secondary.

    See Abreview

  • ICC/IF of U251 cells staining intracellular Nestin filaments with ab22035

  • Lane 1 : Anti-Nestin antibody [10C2] (ab22035) at 1/1000 dilution
    Lane 2 : Anti-Nestin antibody [10C2] (ab22035)

    Lane 1 : U251 cells at 10 µl
    Lane 2 : U251 cells at 20 µl


    Predicted band size : 176 kDa


    Exposure time : 60 seconds

References

This product has been referenced in:
  • Xiong Y  et al. Precursor N-cadherin mediates glial cell line-derived neurotrophic factor-promoted human malignant glioma. Oncotarget 8:24902-24914 (2017). Human . Read more (PubMed: 28212546) »
  • Montali M  et al. Human adult mesangiogenic progenitor cells reveal an early angiogenic potential, which is lost after mesengenic differentiation. Stem Cell Res Ther 8:106 (2017). ICC/IF ; Human . Read more (PubMed: 28464921) »

See all 38 Publications for this product

Customer reviews and Q&As

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (First trimester human placenta)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium citrate
Permeabilization
No
Specification
First trimester human placenta
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Formaldehyde
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Verified customer

Submitted Mar 03 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Pig Cell (bone marrow-mesenchymal stem cell)
Permeabilization
Yes - saponin 0,1%
Specification
bone marrow-mesenchymal stem cell
Blocking step
Protein block serum-free ready to use (DAKO) + saponin 0,1% as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0% · Temperature: 25°C
Fixative
Formaldehyde
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Verified customer

Submitted Aug 23 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Human iPSC derived neural stem cell)
Permeabilization
Yes - 0.1% Triton X-100 in PBS
Specification
Human iPSC derived neural stem cell
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jul 13 2015

Thank you for contacting us.

1. Anti-Nestin antibody [10C2] (ab22035) and Anti-HIF1 alpha antibody [H1alpha67] - BSA and Azide free (ab80544):

Volume: 100 µg at 1 mg/ml

We recommend to start with a concentration of 5 u...

Read More

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...

Read More

Thank you very much for your email. I apologize for the delay in our answer.

I can confirm thatthis antibody can be used with HRP conjugated secondary antibody without any problems.

The note on the datasheet is in reference on how i...

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Application
Flow Cytometry
Sample
Human Cell (Differentiation Human ES cell)
Specification
Differentiation Human ES cell
Preparation
Cell harvesting/tissue preparation method: Cell dissociation buffer
Sample buffer: FACS buffer (0.2%BSA, 0.1%sodium azide in PBS)
Fixation
Formaldehyde
Permeabilization
Yes - 70% methanol (ice cold)
Gating Strategy
FSC vs SSC on negative control sample
Username

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Verified customer

Submitted Jan 09 2012

Thank you for your patience. I have attached the test results from the lab. We performed an immunoblotting experiment on a cell extract of U251 cells. The attached Western blot image clearly indicates a Nestin band with a MW of approximately 250 kDa. T...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (brain tumor cell line)
Specification
brain tumor cell line
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.05% Triton X-100
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jul 29 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (human ES cell derived neurons)
Specification
human ES cell derived neurons
Fixative
Paraformaldehyde
Permeabilization
Yes - See Blocking buffer
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Jun 21 2011

1-10 of 13 Abreviews or Q&A

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