Overview

  • Product name
    Anti-Nestin antibody [Rat 401]
    See all Nestin primary antibodies
  • Description
    Mouse monoclonal [Rat 401] to Nestin
  • Host species
    Mouse
  • Specificity
    The clone number has been updated from (2Q178) to (Rat 401) both clone numbers name the same antibody clone.
  • Tested applications
    Suitable for: ICC/IF, 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 protein (purified) corresponding to Rat Nestin.

  • 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.
Flow Cyt Use at an assay dependent concentration. PubMed: 21092738

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IHC-FoFr Use at an assay dependent concentration. 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 concentration. Predicted molecular weight: 200 kDa. Block with milk or BSA but do not dilute primary antibody in buffer containing milk.

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

  • 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 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

  • ab6142 staining Nestin in mouse brain tissue sections by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with acetone, permeabilized with Triton X-100 in PBS and blocked with 5% BSA for 60 minutes at room temperature. Samples were incubated with primary antibody (1/1000 in PBST) for 12 hours at 4°C. An Alexa Fluor® 488-conjugated goat anti-mouse IgG monoclonal was used as the secondary antibody at a dilution of 1/200.

    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 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

References

This product has been referenced in:
  • Gholinejad M  et al. Adenosine decreases oxidative stress and protects H2O2-treated neural stem cells against apoptosis through decreasing Mst1 expression. Biomed Rep 8:439-446 (2018). Read more (PubMed: 29732147) »
  • Lin GQ  et al. Transplanted human neural precursor cells integrate into the host neural circuit and ameliorate neurological deficits in a mouse model of traumatic brain injury. Neurosci Lett 674:11-17 (2018). IHC . Read more (PubMed: 29501684) »

See all 77 Publications for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

Filter by Ratings

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Brain)
Antigen retrieval step
None
Permeabilization
Yes - TritonX-100
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

Submitted Nov 02 2016

Application
Immunocytochemistry
Sample
Mouse Cell (Mouse neural stem cell)
Permeabilization
Yes - tritonx100
Specification
Mouse neural stem cell
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

Submitted Nov 02 2016

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (brain)
Permeabilization
Yes - Triton
Specification
brain
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Acetone
Username

Abcam user community

Verified customer

Submitted Feb 09 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (adult rat heart tissue)
Permeabilization
Yes - 0.1% Triton
Specification
adult rat heart tissue
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 2% · Temperature: 20°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jan 28 2016

Application
Western blot
Loading amount
400000 cells
Gel Running Conditions
Reduced Denaturing (12%)
Sample
Mouse Cell lysate - whole cell (Mouse Neuroblastoma (Neuro2A))
Specification
Mouse Neuroblastoma (Neuro2A)
Treatment
Serum deprivation (0.1%) for 24 hrs
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 28°C
Username

Abcam user community

Verified customer

Submitted Mar 25 2015

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Apteronotus leptorhynchus Tissue lysate - whole (Brain)
Gel Running Conditions
Reduced Denaturing (4-15%)
Loading amount
50 µg
Specification
Brain
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

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

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

1-10 of 23 Abreviews

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