Anti-Hypophosphorylated Neurofilament H 抗体 [N52] (ab82259)

製品の概要

  • 製品名Anti-Hypophosphorylated Neurofilament H antibody [N52]
  • 製品の詳細
    Mouse monoclonal [N52] to Hypophosphorylated Neurofilament H
  • アプリケーション適用あり: WB, IHC-P, IHC-Fr, IHC-FoFr, Flow Cyt, ICC/IFmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    Other Immunogen Type corresponding to Pig Hypophosphorylated Neurofilament H (C terminal). C-terminal segment of enzymatically dephosphorylated pig Neurofilament 200.

  • 特記事項

    Other applications have not been tested.


    Optimal dilutions should be determined by end users.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab82259 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
WB Use at an assay dependent concentration. Predicted molecular weight: 112 kDa.
IHC-P Use a concentration of 1 - 2 µg/ml.
IHC-Fr Use a concentration of 1 - 2 µg/ml. Acetone fixed.
IHC-FoFr 1/50.
Flow Cyt Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ICC/IF Use at an assay dependent concentration.

ターゲット情報

  • 機能Neurofilaments usually contain three intermediate filament proteins: L, M, and H which are involved in the maintenance of neuronal caliber. NF-H has an important function in mature axons that is not subserved by the two smaller NF proteins.
  • 関連疾患Defects in NEFH are a cause of susceptibility to amyotrophic lateral sclerosis (ALS) [MIM:105400]. ALS is a neurodegenerative disorder affecting upper and lower motor neurons, and resulting in fatal paralysis. Sensory abnormalities are absent. Death usually occurs within 2 to 5 years. The etiology is likely to be multifactorial, involving both genetic and environmental factors.
  • 配列類似性Belongs to the intermediate filament family.
  • 翻訳後修飾There are a number of repeats of the tripeptide K-S-P, NFH is phosphorylated on a number of the serines in this motif. It is thought that phosphorylation of NFH results in the formation of interfilament cross bridges that are important in the maintenance of axonal caliber.
    Phosphorylation seems to play a major role in the functioning of the larger neurofilament polypeptides (NF-M and NF-H), the levels of phosphorylation being altered developmentally and coincident with a change in the neurofilament function.
    Phosphorylated in the Head and Rod regions by the PKC kinase PKN1, leading to inhibit polymerization.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 200 kDa neurofilament protein antibody
    • Nefh antibody
    • Neurofilament heavy polypeptide antibody
    • Neurofilament triplet H protein antibody
    • NF-H antibody
    • NF200 antibody
    • NFH antibody
    • NFH_HUMAN antibody
    see all

Anti-Hypophosphorylated Neurofilament H antibody [N52] 画像

  • ab82259 staining Hypophosphorylated Neurofilament H in Rat nerve tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with formaldehyde, permeablized with 0.2 Triton-X and blocked with 10% serum. The sample was incubated with primary antibody (1/50 in PBS plus 1x Casien) at 25°C for 2 hours. An Alexa Fluor®488-conjugated Goat anti-mouse IgG polyclonal(1/200) was used as the secondary antibody. GFAP was labelled with Texas Red.

    See Abreview

  • ab82259 staining hypophosphorylated Neurofilament H in paraffin-embedded rat brain tissue.
  • ICC/IF image of ab82259 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 (ab82259, 1µg/ml) overnight at +4°C. The secondary antibody (green) was 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 1.43µM.

  • Overlay histogram showing SH-SY5Y cells stained with ab82259 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab82259, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

Anti-Hypophosphorylated Neurofilament H antibody [N52] (ab82259) 使用論文

This product has been referenced in:
  • Zonouzi M  et al. GABAergic regulation of cerebellar NG2 cell development is altered in perinatal white matter injury. Nat Neurosci 18:674-82 (2015). IHC-FoFr ; Mouse . Read more (PubMed: 25821912) »
  • Masliukov PM  et al. Development of nNOS-positive neurons in the rat sensory and sympathetic ganglia. Neuroscience 256C:271-281 (2013). IHC ; Rat . Read more (PubMed: 24161722) »

See all 5 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Penguin Tissue sections (brain)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: tris-EDTA, pH9.0
Permeabilization No
Specification brain
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

投稿 May 02 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Zebra finch Tissue sections (brain)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: tris-EDTA, pH9.0
Permeabilization No
Specification brain
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

投稿 Apr 26 2016

Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (rat motor neuron primary cells from embryo spiral)
Permeabilization Yes - 0.2% triton
Specification rat motor neuron primary cells from embryo spiral
Blocking step Leica Universal IHC blocking as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Aug 06 2015

Application Immunohistochemistry (Frozen sections)
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Sample Rat Tissue sections (spinal cord)
Specification spinal cord
Permeabilization No
Fixative Acetone
Username

Abcam user community

Verified customer

投稿 Mar 24 2015


The antibodies mentioned above are suitable for Rat as well as human samples. ab9018 is yet to be tested in IHC-Fr however as it is tested in IHC-FoFr so it is more likely to work.

The suitable secondary's are for ab4648, ab82259 and ab90...

Read More

Thank you for your enquiry regarding ab82259 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that your customer is having problems with this antibody.

After reading through the detailed proto...

Read More

I am very pleased to hear you would like to test ab5076, ab8049, ab18258, ab58802, ab64581, ab82259 and send us your results. The testing discount doesn't apply to ab4674 since this has already been tested in Rhesus monkey and IHC so we would guarantee...

Read More

Thank you for contacting us.

Yes, the antibody ab82259 can be used to stain sections of paraffin-embedded tissue but the paraffin will need to be removed from the sections. Without paraffin removal, aqueous solutions will not diffuse into th...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Rat Tissue sections (nerve)
Specification nerve
Fixative Formaldehyde
Permeabilization Yes - 0.2% triton
Username

Abcam user community

Verified customer

投稿 May 08 2013

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