重组
RabMAb

Anti-160 kD Neurofilament Medium抗体[EP2460] (ab92539)

概述

性能

应用

Our Abpromise guarantee covers the use of ab92539 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB
IP
IHC-Fr
IHC-P
Flow Cyt
ICC/IF
  • 应用说明
    ICC/IF: 1/100 - 1/250.
    IHC-P: 1/250 - 1/500. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.
    IP: 1/10.
    WB: 1/100000 - 1/500000. Predicted molecular weight: 102 kDa.

    Is unsuitable for Flow Cyt.


    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • 靶标

    • 相关性
      Neurofilaments are the 10nm or intermediate filament proteins found specifically in neurons, and are composed predominantly of three major proteins called neurofilament light (NF-L), neurofilament medium (NF-M) and neurofilament heavy (NF-H). Neurofilament medium runs on SDS-PAGE gels in the range 145-170 kDa, with some variation in different species. Antibodies to this protein are useful to identify neurons and their processes in tissue sections and in tissue culture. Neurofilament medium can also be useful in studies of neurofilament accumulations seen in many neurological diseases, such as Lou Gehrig's disease or Alzheimer's disease.
    • 细胞定位
      Cytoplasm
    • 数据库链接
    • 别名
      • 150kDa medium antibody
      • NEF3 antibody
      • NEFM antibody
      • Neurofilament 3 antibody
      • NF160 antibody
      • NFM antibody
      see all

    图片

    • ab92539 stained in SHSY5Y cells. Cells were fixed with 100% methanol (5 min), permeabilized in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h.The cells were then incubated with the antibody ab92539 at 0.5µg/ml and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150087 (colored red) and ab150177 (colored green) used at 1 ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature.

      This product gave a positive signal in 4% formaldehyde (10 min) fixed SHSY5Y cells under the same testing conditions.

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    • IHC image of Neurofilament 160 kDa staining in a section of frozen normal human cerebral cortex*.

      The section was fixed using 10% formaldehyde in 1XPBS for 10 minutes. No antigen retrieval step was performed prior to staining. Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 1% (w/v) BSA for 1h at room temperature.  The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab92539 at 1/500 dilution. The secondary antibody was ab150081 (colored green) used at 1 ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature.The section was then mounted using Fluoromount®.

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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

      *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

    • ab92539, at 1/250 dilution, staining 160 kD Neurofilament Medium in formalin-fixed, paraffin-embedded Human normal brain tissue by immunohistochemistry.
    • All lanes : Anti-160 kD Neurofilament Medium antibody [EP2460] (ab92539) at 1/100000 dilution

      Lane 1 : 293T cell lysate
      Lane 2 : SH-SY5Y cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      HRP labelled Goat anti-Rabbit IgG at 1/2000 dilution

      Predicted band size : 102 kDa
    • ab92539, at 1/250 dilution, staining 160 kD Neurofilament Medium in formalin-fixed, paraffin-embedded Human glioma tissue by immunohistochemistry.
    • ab92539 showing negative staining in Normal breast tissue.

    • ab92539 showing negative staining in Normal colon tissue.

    • ab92539 showing negative staining in Normal tonsil tissue.

    • ab92539 showing negative staining in Normal liver tissue.

    • ab92539 showing negative staining in Hepatocellular carcinoma tissue.

    • ab92539 showing negative staining in Normal colon tissue.

    • ab92539 showing negative staining in Normal liver tissue.

    文献

    This product has been referenced in:
    • Foster JW  et al. Low-glucose enhances keratocyte-characteristic phenotype from corneal stromal cells in serum-free conditions. Sci Rep 5:10839 (2015). ICC/IF ; Human . Read more (PubMed: 26039975) »
    • Rettig I  et al. Selective inhibition of HDAC8 decreases neuroblastoma growth in vitro and in vivo and enhances retinoic acid-mediated differentiation. Cell Death Dis 6:e1657 (2015). Flow Cyt ; Human . Read more (PubMed: 25695609) »

    See all 2 Publications for this product

    客户评价及客户问答

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Immunohistochemistry (Frozen sections)
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
    Sample
    Human Tissue sections (Human Skin from dermal and epidermal cells)
    Specification
    Human Skin from dermal and epidermal cells
    Permeabilization
    Yes - 0.01% Triton X
    Fixative
    Acetone
    Username

    Abcam user community

    Verified customer

    提交于 Nov 21 2012

    Thank you for contacting us.

    This target should only be expressed in neuronal tissues. Therefore any non-neuronal tissues should serve as a negative control. We have shown negative staining in normal breast tissue, normal colon tissue, nor...

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    Thank you for your phone call.

    I am sorry to hear that both antibodies do not work as expected.


    As for ab92539, I am sending you - as we discussed - the testing discount code for having tried it in IHC-Fr. After submitting you...

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

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