製品の概要

  • 製品名
    Anti-160 kD Neurofilament Medium antibody
    160 kD Neurofilament Medium 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to 160 kD Neurofilament Medium
  • アプリケーション
    適用あり: IHC-FoFr, IHC-P, WB, ICC/IFmore details
  • 種交差性
    交差種: Mouse, Rat, Human
    交差が予測される動物種: Cow, Rhesus monkey
  • 免疫原

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

    (Peptide available as ab66467.)

  • ポジティブ・コントロール
    • This antibody gave a positive signal in the following tissue lysates; Mouse Spinal Cord; Rat Spinal Cord; Mouse Cerebellum; Rat Cerebellum; Human Spinal Cord.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

ターゲット情報

  • 関連性
    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.
  • 細胞内局在
    Cytoplasm
  • 参照データベース
  • 別名
    • 150kDa medium antibody
    • NEF3 antibody
    • NEFM antibody
    • Neurofilament 3 antibody
    • NF160 antibody
    • NFM antibody
    see all

画像

  • 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

  • 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 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.

参考文献

This product has been referenced in:
  • Shah B  et al. C3G/Rapgef1 Is Required in Multipolar Neurons for the Transition to a Bipolar Morphology during Cortical Development. PLoS One 11:e0154174 (2016). Read more (PubMed: 27111087) »
  • Thaung HP  et al. Increased Efferent Cardiac Sympathetic Nerve Activity and Defective Intrinsic Heart Rate Regulation in Type 2 Diabetes. Diabetes 64:2944-56 (2015). Read more (PubMed: 25784543) »

See all 3 Publications for this product

レビューと Q&A

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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 (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Citrate buffer pH6
Permeabilization
No
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% Normal buffered formalin
Username

Abcam user community

Verified customer

投稿 Dec 05 2016

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)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Citrate buffer pH6
Permeabilization
No
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% Normal buffered formalin
Username

Abcam user community

Verified customer

投稿 Dec 05 2016

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

Abcam user community

Verified customer

投稿 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

投稿 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

投稿 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

投稿 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

投稿 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

投稿 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

投稿 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

投稿 Nov 17 2010

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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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