Anti-160 kD Neurofilament Medium antibody (ab64300)

Overview

  • Product nameAnti-160 kD Neurofilament Medium antibody
    See all 160 kD Neurofilament Medium primary antibodies
  • Description
    Rabbit polyclonal to 160 kD Neurofilament Medium
  • Tested applicationsSuitable for: IHC-FoFr, IHC-P, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Cow, Rhesus monkey
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 800 to the C-terminus of Rat 160 kD Neurofilament Medium.

    (Peptide available as ab66467.)

  • Positive control
    • This antibody gave a positive signal in the following tissue lysates; Mouse Spinal Cord; Rat Spinal Cord; Mouse Cerebellum; Rat Cerebellum; Human Spinal Cord.

Properties

Applications

Our Abpromise guarantee covers the use of ab64300 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
IHC-FoFr 1/100.
IHC-P 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 145 kDa (predicted molecular weight: 96 kDa).
ICC/IF Use a concentration of 1 µg/ml.

Target

  • RelevanceNeurofilaments 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 localizationCytoplasm
  • Database links
  • Alternative names
    • 150kDa medium antibody
    • NEF3 antibody
    • NEFM antibody
    • Neurofilament 3 antibody
    • NF160 antibody
    • NFM antibody
    see all

Anti-160 kD Neurofilament Medium antibody images

  • Anti-160 kD Neurofilament Medium antibody (ab64300) at 1 µg/ml + Human spinal cord tissue lysate - total protein (ab29188) at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Predicted band size : 96 kDa
    Observed band size : 166 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 50 kDa,53 kDa. We are unsure as to the identity of these extra bands.
  • All lanes : Anti-160 kD Neurofilament Medium antibody (ab64300) at 1 µg/ml

    Lane 1 : Spinal Cord (Mouse) Tissue Lysate
    Lane 2 : Spinal Cord (Rat) Tissue Lysate
    Lane 3 : Cerebellum Mouse Tissue Lysate
    Lane 4 : Cerebellum Rat Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size : 96 kDa
    Observed band size : 145 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 50 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 4 minutes
  • ab64300 staining rat cerebellum sections by IHC-P. The tissue was fixed with formaldehyde and a heat mediated antigen retrival step was performed with citric acid pH 6. Blocking of the sample was done with 1% BSA for 10 minutes at 21°C, followed by staining with ab64300 at 1/2000 in TBS/BSA/azide for 16h at 21°C. A biotinylated goat anti-rabbit polyclonal antibody at 1/200 was used as the secondary antibody.

    See Abreview

  • ab64300 staining mouse cerebellum sections by IHC-P. The tissue was fixed with formaldehyde and a heat mediated antigen retrival step was performed with citric acid pH 6. Blocking of the sample was done with 1% BSA for 10 minutes at 21°C, followed by staining with ab64300 at 1/2000 in TBS/BSA/azide for 16h at 21°C. A biotinylated goat anti-rabbit polyclonal antibody at 1/200 was used as the secondary antibody.

    See Abreview

  • ab64300 (1/50) staining 160 kD Neurofilament Medium in paraffin-embedded Human brain tissue. Tissue underwent fixation in formaldehyde, peroxidase blocking, protein blocking and heat mediated antigen retrieval. The secondary antibody was goat anti rabbit conjugated to HRP. For further experimental details please refer to abreview.

    See Abreview

  • IHC-FoFr image of 160kda Neurofilament Medium staining on mouse brain sections using ab64300(1:100). The sections used came from animals perfused fixed with Paraformaldehyde 4% with 15% of a solution of saturated picric acid, in phosphate buffer 0.1M. Following postfixation in the same fixative overnight, the brains were cryoprotected in sucrose 30% overnight. Brains were then cut using a cryostat and the immunostainings were performed using the ‘free floating’ technique.

    See Abreview

  • ICC/IF image of ab64300 stained PC12 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 (ab64300, 5µg/ml) 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.

References for Anti-160 kD Neurofilament Medium antibody (ab64300)

This product has been referenced in:
  • Khan RS  et al. SIRT1 Activating compounds reduce oxidative stress mediated neuronal loss in viral induced CNS demyelinating disease. Acta Neuropathol Commun 2:3 (2014). IHC . Read more (PubMed: 24383546) »

See 1 Publication for this product

Product Wall

Application IHC - Wholemount
Sample Mouse Tissue (Diaphragm)
Specification Diaphragm
Username

Abcam user community

Verified customer

Submitted Oct 21 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (embryonic brain cortex and hippocampus)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10mM sodium citrate buffer pH 6
Specification embryonic brain cortex and hippocampus
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative Carnoys fixative
Username

Dr. Bhavin Shah

Verified customer

Submitted Oct 19 2015

Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (Cryopreserved dorsal root ganglion neurons (QBMce)
Specification Cryopreserved dorsal root ganglion neurons (QBMce
Permeabilization No
Fixative paraformaldehyde with picric acid
Username

Ms. Babben Tinner

Verified customer

Submitted Feb 26 2015

Application Immunohistochemistry (Frozen sections)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Sample Zebrafish Tissue sections (Retina, Neurons)
Specification Retina, Neurons
Permeabilization Yes - Triton X
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Aug 29 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Mouse Tissue sections (Brain)
Specification Brain
Fixative Paraformaldehyde
Permeabilization No
Username

Dr. Sophie Pezet

Verified customer

Submitted Jun 08 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Cerebellum)
Specification Cerebellum
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Username

Mr. Carl Hobbs

Verified customer

Submitted Mar 20 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Rat Tissue sections (Cerebellum)
Specification Cerebellum
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Username

Mr. Carl Hobbs

Verified customer

Submitted Mar 20 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Brain)
Specification Brain
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Dako Target Retreival Solution
Permeabilization No
Blocking step 5 minutes of peroxidase block then 10 minutes of protein block. These are ready-to-use reagents purchased from Dako as blocking agent for 15 minute(s) · Concentration: 100% · Temperature: 20°C
Username

Mr. Antibody Solutions

Verified customer

Submitted Nov 17 2010

Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Mouse Tissue sections (Brain sections)
Specification Brain sections
Fixative Paraformaldehyde
Antigen retrieval step None
Permeabilization No
Username

Dr. Sophie Pezet

Verified customer

Submitted Aug 12 2009

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