Overview

  • Product name
    Anti-Nestin antibody [SP103]
    See all Nestin primary antibodies
  • Description
    Rabbit monoclonal [SP103] to Nestin
  • Tested applications
    Suitable for: ICC/IF, Flow Cyt, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide corresponding to the C terminus of Human Nestin.

  • Positive control
    • HeLa cell lysate and Human kidney tissue. SKNSH cell line
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab105389 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 1/200.
Flow Cyt 1/50.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB 1/100. Predicted molecular weight: 177 kDa.
IHC-P 1/100.

Antigen retrieval: Boil tissue section in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at room temperature for 20 min.

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

Anti-Nestin antibody [SP103] images

  • Anti-Nestin antibody [SP103] (ab105389) at 1/100 dilution + HeLa cell lysate

    Predicted band size : 177 kDa
  • Immunohistochemical staining of human kidney with ab105389 at 1/100.

  • ab105389 staining Nestin in human melanoma tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
    Tissue was fixed in formaldehyde and a heat mediated antigen retrieval step was performed using Tris EDTA pH 9.0. Samples were then permeabilized using 0.25% Triton X-100, blocked, then incubated with ab105389 at a 1/100 dilution for 18 hours at 4°C. The secondary used was an Alexa-Fluor 594 (red) conjugated donkey anti-rabbit polyclonal used at a 1/100 dilution. (Green) tumor cells, (blue) DAPI.

    See Abreview

  • ICC/IF image of ab105389 stained SKNSH cells. The cells were 4% formaldehyde fixed (10 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 (ab105389, 1/200 dilution) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.
  • Overlay histogram showing SH-SY5Y cells stained with ab105389 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab105389, 1/50 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References for Anti-Nestin antibody [SP103] (ab105389)

This product has been referenced in:
  • Wenger A  et al. Stem cell cultures derived from pediatric brain tumors accurately model the originating tumors. Oncotarget 8:18626-18639 (2017). IHC-P ; Mouse . Read more (PubMed: 28148893) »
  • Malchenko S  et al. Stabilization of HIF-1a and HIF-2a, up-regulation of MYCC and accumulation of stabilized p53 constitute hallmarks of CNS-PNET animal model. PLoS One 12:e0173106 (2017). IHC-P ; Human . Read more (PubMed: 28249000) »

See all 8 Publications for this product

Product Wall

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Dako protein block as blocking agent for 10 minute(s) · Concentration: .25% · Temperature: 22°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate
Sample
Dog Tissue sections (Fetal tissue)
Specification
Fetal tissue
Permeabilization
No
Fixative
Formaldehyde
Username

Ms. Erica Sloma

Verified customer

Submitted Aug 21 2014

Application
Western blot
Loading amount
5 µg
Gel Running Conditions
Reduced Denaturing (10%)
Sample
Human Cell lysate - other (undifferentiated human iPS cells)
Specification
undifferentiated human iPS cells
Blocking step
5% milk in TBST as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Dec 18 2013

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
1% serum, 0.1% triton, 0.1% BSA in PBS as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: rt°C
Sample
Human Cell (neural progenitor cell derived from human iPSCs)
Specification
neural progenitor cell derived from human iPSCs
Permeabilization
Yes - (see blocking buffer)
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Nov 22 2013

Thank you for contacting us. I am sorry to hear that this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. Read More

Thank you for contacting us.

Unfortunately, alignments of the immunogen with mouse show very poor homology. As such, we cannot guarantee or recommend this product for use in this species. We do offer a number of other anti-Nestin antibodie...

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Thank you for contacting us.

The concentration of lot in question is XXXug/ml (or XX mg/ml).

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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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
Human Tissue sections (kidney)
Specification
kidney
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: EDTA pH9, 20 min, 100C
Permeabilization
No
Username

Abcam user community

Verified customer

Submitted Nov 22 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Melanoma)
Specification
Melanoma
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris EDTA pH9.0
Permeabilization
Yes - 0.25% Triton X-100
Blocking step
DakoCytomation X0909 as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: RT°C
Username

Hongwei Shao

Verified customer

Submitted Nov 09 2011

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

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