Overview

  • Product name

    Anti-Nestin antibody [EPR1301(2)]
    See all Nestin primary antibodies
  • Description

    Rabbit monoclonal [EPR1301(2)] to Nestin
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, Flow Cyt, IHC-Pmore details
    Unsuitable for: IP or WB
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Nestin aa 550-650 (Cysteine residue). The exact sequence is proprietary.
    Database link: P48681

  • Positive control

    • IHC-P: Human kidney, spleen and melanoma tissues. ICC/IF: This antibody gave a positive result when used in the following methanol fixed cell lines: SKNSH, U-87 MG FC: SH-SY5Y
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab176571 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 1/50 - 1/100.

For unpurified use at 1/50 - 1/100.

Flow Cyt 1/160.
IHC-P 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

The use of an HRP/AP polymerized secondary antibody is recommended.

See IHC antigen retrieval protocols.

For unpurified use at 1/250 - 1/500.

  • Application notes
    Is unsuitable for IP or WB.
  • 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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human spleen tissue sections labeling Nestin with purified ab176571 at 1:1000 dilution (1.65 µg/ml). Heat mediated antigen retrieval was performed Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human kidney tissue sections labeling Nestin with purified ab176571 at 1:1000 dilution (1.65 µg/ml). Heat mediated antigen retrieval was performed Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • Immunocytochemistry/ Immunofluorescence analysis of U-87 MG (Human glioblastoma-astrocytoma epithelial cell) cells labeling Nestin with purified ab176571 at 1:150 dilution (10 µg/ml). Cells were fixed in 100% Methanol and permeabilized with None. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
    • ICC/IF image of ab176571 stained SK-N-SH (Human neuroblastoma cell line) cells.

      The cells were fixed in 100% methanol (5 min) and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab176571 at 5 µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti-rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43 µM for 1hour at room temperature.

    • Immunohistochemical analysis of paraffin-embedded human melanoma tissue labeling Nestin using ab176571 at a 1/250 dilution.

    • Flow Cytometry analysis of SH-SY5Y (human neuroblastoma) cells labeling Nestin with purified ab176571 at 1/160 dilution (10 µg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

    • Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Nestin using ab176571 at a 1/250 dilution.

    References

    This product has been referenced in:

    • Chen C  et al. Neuropathy correlated with imbalanced Foxp3/IL-17 in bone marrow microenvironment of patients with acute myeloid leukemia. Oncotarget 7:24455-65 (2016). IHC-P ; Human . Read more (PubMed: 27016413) »
    See 1 Publication for this product

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