Overview

  • Product nameAnti-Nestin antibody [SP103]See all Nestin primary antibodies ...
  • Description
    Rabbit monoclonal [SP103] to Nestin
  • Tested applicationsICC/IF, Flow Cyt, WB, IHC-P more 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

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 Notes
ICC/IF 1/200.
Flow Cyt 1/50.
WB 1/100. Predicted molecular weight: 177 kDa.
IHC-P 1/100.

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.
  • Target information above from: UniProt accession P48681 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    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
  • ab105389, at a 1/100 dilution, staining Nestin in formalin fixed, paraffin embedded Human kidney tissue by Immunohistochemistry.
  • 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:
  • Bury M  et al. Ophiobolin A induces paraptosis-like cell death in human glioblastoma cells by decreasing BKCa channel activity. Cell Death Dis 4:e561 (2013). WB ; Human . Read more (PubMed: 23538442) »

See 1 Publication for this product

Product Wall

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
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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
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Abcam user community

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

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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
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Submitted Nov 22 2011

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
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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"