Overview

  • Product name

    Anti-160 kD Neurofilament Medium antibody [NF-09]
    See all 160 kD Neurofilament Medium primary antibodies
  • Description

    Mouse monoclonal [NF-09] to 160 kD Neurofilament Medium
  • Host species

    Mouse
  • Specificity

    This antibody reacts with both phosphorylated and non-phosphorylated forms of medium neurofilament protein (160 kDa) of various species.
  • Tested applications

    Suitable for: ELISA, IHC-Fr, ICC, IHC-P, WB, IHC (PFA fixed), ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Cow, Cat, Human, Pig
    Predicted to work with: a wide range of other species, Mammals
  • Immunogen

    Pellet of pig brain cold stable proteins after depolymerization of microtubules.

  • Positive control

    • WB: HEK-293 and A549 whole cell lysates ICC/IF: Dental pulp stem, Neuro2A, PC12 and human nerve cells

Properties

Applications

Our Abpromise guarantee covers the use of ab7794 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
ELISA Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
WB 1/1000.
IHC (PFA fixed) 1/500.
ICC/IF Use at an assay dependent concentration.

Target

  • Relevance

    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.
  • Cellular localization

    Cytoplasm
  • Database links

  • Alternative names

    • 150kDa medium antibody
    • NEF3 antibody
    • NEFM antibody
    • Neurofilament 3 antibody
    • NF160 antibody
    • NFM antibody
    see all

Images

  • All lanes : Anti-160 kD Neurofilament Medium antibody [NF-09] (ab7794) at 1/1000 dilution

    Lane 1 : Wild-type HEK-293 whole cell lysate
    Lane 2 : NEFM knockout HEK-293 whole cell lysate
    Lane 3 : A549 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Observed band size: 150 kDa
    why is the actual band size different from the predicted?



    Lanes 1 - 4: Merged signal (red and green). Green - ab7794 observed at 150 kDa. Red - loading control, ab52866, observed at 50 kDa.

    ab7794 was shown to specifically react with NEFM (Neurofilament) in wild-type HEK-293 cells as signal was lost in NEFM knockout cells. Wild-type and NEFM knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% NF Milk. Ab7794 and ab52866 (Rabbit anti alpha Tubulin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

     

  • ab7794 staining 160 kD Neurofilament Medium (red) in Human nerve surrounding common hepatic artery tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/500 in Da Vinci Green diluent) for 16 hours at 4°C. An undiluted HRP-conjugated mouse polymer was used as the secondary antibody. Tyrosin Hydroxylase was stainded green with ab112

    See Abreview

  • Western blotting analysis of neurofilament medium protein in porcine brain lysate (reducing conditions) by mouse monoclonal NF-09.

  • Immunofluorescence analysis of 1 month neuronal-differentiated dental pulp stem cells, staining 160 kD Neurofilament Medium with ab7794.

    Cells were fixed with paraformaldehyde and incubated with primary antibody (1/600). A FITC-conjugated anti-mouse IgG (1/375) was used as the secondary antibody.
  • ab7794 staining Neurofilament medium protein in mouse Neuro2A cells by ICC/IF.
  • ICC/IF image of ab7794 stained PC12 cells. The cells were 100% methanol fixed (5 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 (ab7794, 1æg/ml) overnight at +4øC. The secondary antibody (green)ÿwas Alexa Fluor© 488 goat anti-mouse IgG (H+L) used at a 1/1000 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 1.43æM.

References

This product has been referenced in:

  • Jung W  et al. Wire Electrodes Embedded in Artificial Conduit for Long-term Monitoring of the Peripheral Nerve Signal. Micromachines (Basel) 10:N/A (2019). Read more (PubMed: 30871203) »
  • Huang CW  et al. Multipotent vascular stem cells contribute to neurovascular regeneration of peripheral nerve. Stem Cell Res Ther 10:234 (2019). Read more (PubMed: 31376835) »
See all 26 Publications for this product

Customer reviews and Q&As

Filter by Application

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1-2 of 2 Abreviews

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Nerve surrounding common hepatic artery)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer pH 6.0
Permeabilization
No
Specification
Nerve surrounding common hepatic artery
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jan 14 2017

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (nerve)
Specification
nerve
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.3% TritonX-100
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jun 23 2010

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