Overview

  • Product name

    Anti-Nestin antibody [rat-401] (HRP)
    See all Nestin primary antibodies
  • Description

    Mouse monoclonal [rat-401] to Nestin (HRP)
  • Host species

    Mouse
  • Conjugation

    HRP
  • Tested applications

    Suitable for: WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat
  • Immunogen

    Full length native protein (purified) corresponding to Human Nestin. Nestin purified from embryonic rat spinal cord.

  • Positive control

    • WB: MEF1 whole cell lysate. IHC-P: FFPE rat 6 week brain (SVZ) tissue sections.

Properties

Applications

Our Abpromise guarantee covers the use of ab196694 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
WB 1/2000. Detects a band of approximately 170 kDa (predicted molecular weight: 177 kDa).
IHC-P 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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

  • Anti-Nestin antibody [rat-401] (HRP) (ab196694) at 1/2000 dilution + MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 177 kDa
    Observed band size: 170 kDa
    why is the actual band size different from the predicted?


    Exposure time: 20 minutes


    This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab196694 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

  • IHC image of Nestin staining in a section of formalin-fixed paraffin-embedded rat 6 week brain (SVZ), performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab196694, 1/50 dilution, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

     

References

ab196694 has not yet been referenced specifically in any publications.

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