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使用敲除细胞株进行验证

Anti-160 kD Neurofilament Medium抗体[NF-09] - Neuronal Marker (ab7794)

  • Datasheet
  • SDS
Reviews (5)Q&A (1)References (47)

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Western blot - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)
  • Immunocytochemistry/ Immunofluorescence - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)
  • Western blot - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)
  • Western blot - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)
  • Immunocytochemistry/ Immunofluorescence - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)
  • Immunocytochemistry/ Immunofluorescence - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)

Key features and details

  • Mouse monoclonal [NF-09] to 160 kD Neurofilament Medium - Neuronal Marker
  • Suitable for: WB, ELISA, ICC/IF, IHC-Fr, IHC-P, IHC (PFA fixed)
  • Knockout validated
  • Reacts with: Mouse, Rat, Cow, Cat, Human, Pig
  • Isotype: IgG2a

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概述

  • 产品名称

    Anti-160 kD Neurofilament Medium抗体[NF-09] - Neuronal Marker
    参阅全部 160 kD Neurofilament Medium 一抗
  • 描述

    小鼠单克隆抗体[NF-09] to 160 kD Neurofilament Medium - Neuronal Marker
  • 宿主

    Mouse
  • 特异性

    This antibody reacts with both phosphorylated and non-phosphorylated forms of medium neurofilament protein (160 kDa) of various species.
  • 经测试应用

    适用于: WB, ELISA, ICC/IF, IHC-Fr, IHC-P, IHC (PFA fixed)more details
  • 种属反应性

    与反应: Mouse, Rat, Cow, Cat, Human, Pig
    预测可用于: a wide range of other species, Mammals
  • 免疫原

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

  • 阳性对照

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

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

性能

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • 存储溶液

    pH: 7.40
    Preservative: 0.097% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • 纯度

    Protein A purified
  • 克隆

    单克隆
  • 克隆编号

    NF-09
  • 同种型

    IgG2a
  • 轻链类型

    unknown
  • 研究领域

    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Alzheimer's disease
    • Other
    • Neuroscience
    • Cell Adhesion Proteins
    • Cytoskeletal Proteins
    • Intermediate Filaments
    • Neuroscience
    • Cell Type Marker
    • Neuron marker
    • Soma marker
    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Intermediate Filaments
    • Class IV
    • Neurofilaments
    • Stem Cells
    • Lineage Markers
    • Ectoderm
    • Developmental Biology
    • Lineage specification
    • Ectoderm
    • Neuroscience
    • Development

相关产品

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
  • Isotype control

    • Mouse IgG2a, kappa monoclonal [MG2a-53] - Isotype control (ab18415)
  • KO cell lines

    • Human NEFM (160 kD Neurofilament Medium) knockout HEK-293T cell line (ab266741)
  • KO cell lysates

    • Human NEFM (160 kD Neurofilament Medium) knockout HEK-293T cell lysate (ab257103)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab7794于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
WB
Use a concentration of 1 - 2 µg/ml.
ELISA
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.
IHC-Fr (1)
Use at an assay dependent concentration.
IHC-P (1)
Use at an assay dependent concentration.
IHC (PFA fixed)
Use at an assay dependent concentration.
说明
WB
Use a concentration of 1 - 2 µg/ml.
ELISA
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.
IHC-Fr
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration.
IHC (PFA fixed)
Use at an assay dependent concentration.

靶标

  • 功能

    Neurofilaments usually contain three intermediate filament proteins: L, M, and H which are involved in the maintenance of neuronal caliber.
  • 序列相似性

    Belongs to the intermediate filament family.
  • 翻译后修饰

    There are a number of repeats of the tripeptide K-S-P, NFM is phosphorylated on a number of the serines in this motif. It is thought that phosphorylation of NFM 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 coincidentally with a change in the neurofilament function.
    Phosphorylated in the head and rod regions by the PKC kinase PKN1, leading to the inhibition of polymerization.
  • Target information above from: UniProt accession P07197 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 数据库链接

    • Entrez Gene: 281347 Cow
    • Entrez Gene: 4741 Human
    • Entrez Gene: 18040 Mouse
    • Entrez Gene: 24588 Rat
    • Omim: 162250 Human
    • SwissProt: O77788 Cow
    • SwissProt: P07197 Human
    • SwissProt: P08553 Mouse
    • SwissProt: P12839 Rat
    • Unigene: 458657 Human
    • Unigene: 730072 Human
    • Unigene: 242832 Mouse
    • Unigene: 390700 Mouse
    • Unigene: 10971 Rat
    see all
  • 别名

    • 150kDa medium antibody
    • 160 kDa neurofilament protein antibody
    • NEF3 antibody
    • NEFM antibody
    • Neurofilament 3 antibody
    • Neurofilament medium polypeptide antibody
    • Neurofilament triplet M protein antibody
    • NF-M antibody
    • NF160 antibody
    • NFM antibody
    • NFM_HUMAN antibody
    see all

图片

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

    Lane 1 : Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
    Lane 2 : NEFM knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
    Lane 3 : A549 (Human lung carcinoma cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) at 1/10000 dilution

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



    Lanes 1-3: Merged signal (red and green). Green - ab7794 observed at 150 kDa. Red - loading control ab181602 observed at 36 kDa.

     ab7794 Anti-160 kD Neurofilament Medium antibody [NF-09]  was shown to specifically react with 160 kD Neurofilament Medium in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266741 (knockout cell lysate ab257103) was used. Wild-type and 160 kD Neurofilament Medium knockout samples were subjected to SDS-PAGE. ab7794 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)
    Immunocytochemistry/ Immunofluorescence - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)Image from Ferro F et al., PLoS One. 2012;7(7):e41774. Epub 2012 Jul 23. Fig 3.; doi:10.1371/journal.pone.0041774; July 23, 2012, PLoS ONE 7(7): e41774.
    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.
  • Western blot - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)
    Western blot - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)
    All lanes : Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (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.

     

  • Western blot - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)
    Western blot - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)

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

  • Immunocytochemistry/ Immunofluorescence - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)
    Immunocytochemistry/ Immunofluorescence - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)
    ab7794 staining Neurofilament medium protein in mouse Neuro2A cells by ICC/IF.
  • Immunocytochemistry/ Immunofluorescence - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)
    Immunocytochemistry/ Immunofluorescence - Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker (ab7794)
    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.

实验方案

  • IHC-FoFr protocol

Click here to view the general protocols

数据表及文件

  • SDS download

  • Datasheet download

    Download

文献 (47)

发表研究结果有使用 ab7794?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab7794 被引用在 47 文献中.

  • Zhang W  et al. Autophagic Schwann cells promote perineural invasion mediated by the NGF/ATG7 paracrine pathway in pancreatic cancer. J Exp Clin Cancer Res 41:48 (2022). PubMed: 35109895
  • Pham VM & Thakor N Insulin enhances neurite extension and myelination of diabetic neuropathy neurons. Korean J Pain 35:160-172 (2022). PubMed: 35354679
  • Company V  et al. Wnt1 Role in the Development of the Habenula and the Fasciculus Retroflexus. Front Cell Dev Biol 9:755729 (2021). PubMed: 34722541
  • Zhao J  et al. Cell-fate transition and determination analysis of mouse male germ cells throughout development. Nat Commun 12:6839 (2021). PubMed: 34824237
  • Fan J  et al. Reinvestigating Tumor-Ventricle Relationship of Craniopharyngiomas With Predominantly Ventricular Involvement: An Endoscopic Endonasal Series Based on Histopathological Assessment. Front Oncol 11:740410 (2021). PubMed: 34926255
View all Publications for this product

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1-6 of 6 Abreviews or Q&A

Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Dog Tissue sections (Brain)
Antigen retrieval step
None
Permeabilization
Yes - 0.1%TritonX100
Specification
Brain
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 24°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Jun 25 2020

Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Peripheral nerve)
Antigen retrieval step
None
Permeabilization
Yes - TritonX-100
Specification
Peripheral nerve
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 24°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 May 27 2020

Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Horse Tissue sections (Peripheral nerve)
Antigen retrieval step
None
Permeabilization
Yes - TritonX-100
Specification
Peripheral nerve
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 24°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 May 22 2020

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Jan 14 2017

Immunohistochemistry (Frozen sections) abreview for Anti-160 kD Neurofilament Medium antibody [NF-09] - Neuronal Marker

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (nerve)
Permeabilization
Yes - 0.3% TritonX-100
Specification
nerve
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Jun 23 2010

Question

I am interested in purchasing a positive control for the intermediate neurofilament (160) antibody for Western Blot usage.

Read More

Abcam community

Verified customer

Asked on Jun 25 2004

Answer

The originator of ab7794 uses cell brain lysate (pig or mouse) as a positive control in Western blots. If you have any more questions, please contact us again.

Read More

Abcam Scientific Support

回复于 Jun 29 2004

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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