Anti-68kDa Neurofilament/NF-L 抗体 [DA2] (ab7255)

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Key features and details

  • Mouse monoclonal [DA2] to 68kDa Neurofilament/NF-L
  • Suitable for: Flow Cyt, ICC/IF, WB, IHC-P
  • Reacts with: Human
  • Isotype: IgG1

製品の概要

  • 製品名

    Anti-68kDa Neurofilament/NF-L antibody [DA2]
    68kDa Neurofilament/NF-L 一次抗体 製品一覧

  • 製品の詳細

    Mouse monoclonal [DA2] to 68kDa Neurofilament/NF-L

  • 由来種

    Mouse

  • 特異性

    Specifically recognizes the light neurofilament subunit or 68kDa Neurofilament/NF-L.

  • アプリケーション

    適用あり: Flow Cyt, ICC/IF, WB, IHC-Pmore details

  • 種交差性

    交差種: Human
    交差が予測される動物種: Bird, Mammals

  • 免疫原

    Full length native protein (purified) corresponding to Pig 68kDa Neurofilament/NF-L. A preparation of enzymatically dephosphorylated pig neurofilaments including NF-L, NF-M and NF-H. Screening was by ELISA on the immunogen followed by immunofluorescence microscopy.

  • エピトープ

    The epitope for this antibody is not in the N-terminus; however exactly where it is located is not known. It is thought to be somewhere within the central alpha helical "rod" region of the molecule.

  • 特記事項

    This product is now provided in a purified format. Lot number GR294347 is still unpurified all other lot numbes are now purified. Dilutions may needed to be ajusted accordingly. 

    Previously labelled as 68kDa Neurofilament. 

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab7255 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
Flow Cyt 1/100.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 
ICC/IF 1/1000.

For unpurifed use at : 1/100 - 1/500
WB 1/5000 - 1/10000.
IHC-P 1/1000.

(works on mildly formalin fixed sections)

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.

  • 関連疾患

    Defects in NEFL are the cause of Charcot-Marie-Tooth disease type 1F (CMT1F) [MIM:607734]. CMT1F is a form of Charcot-Marie-Tooth disease, the most common inherited disorder of the peripheral nervous system. Charcot-Marie-Tooth disease is classified in two main groups on the basis of electrophysiologic properties and histopathology: primary peripheral demyelinating neuropathy or CMT1, and primary peripheral axonal neuropathy or CMT2. Neuropathies of the CMT1 group are characterized by severely reduced nerve conduction velocities (less than 38 m/sec), segmental demyelination and remyelination with onion bulb formations on nerve biopsy, slowly progressive distal muscle atrophy and weakness, absent deep tendon reflexes, and hollow feet. CMT1F is characterized by onset in infancy or childhood (range 1 to 13 years).
    Defects in NEFL are the cause of Charcot-Marie-Tooth disease type 2E (CMT2E) [MIM:607684]. CMT2E is an autosomal dominant form of Charcot-Marie-Tooth disease type 2. Neuropathies of the CMT2 group are characterized by signs of axonal regeneration in the absence of obvious myelin alterations, normal or slightly reduced nerve conduction velocities, and progressive distal muscle weakness and atrophy.

  • 配列類似性

    Belongs to the intermediate filament family.

  • ドメイン

    The extra mass and high charge density that distinguish the neurofilament proteins from all other intermediate filament proteins are due to the tailpiece extensions. This region may form a charged scaffolding structure suitable for interaction with other neuronal components or ions.

  • 翻訳後修飾

    O-glycosylated.
    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

      • 68 kDa neurofilament protein antibody
      • 68kDa Neurofilament antibody
      • 68kDa neurofilament protein antibody
      • CMT1F antibody
      • CMT2E antibody
      • FLJ53642 antibody
      • Light molecular weight neurofilament protein antibody
      • NEFL antibody
      • Neurofilament light antibody
      • Neurofilament light polypeptide 68kDa antibody
      • Neurofilament light polypeptide antibody
      • Neurofilament protein, light chain antibody
      • Neurofilament subunit NF L antibody
      • Neurofilament triplet L protein antibody
      • NF-L antibody
      • NF68 antibody
      • NFL antibody
      • NFL_HUMAN antibody
      see all

    画像

    • Western blot - Anti-68kDa Neurofilament/NF-L antibody [DA2] (ab7255)
      Western blot - Anti-68kDa Neurofilament/NF-L antibody [DA2] (ab7255)
      All lanes : Anti-68kDa Neurofilament/NF-L antibody [DA2] (ab7255) at 1/5000 dilution

      Lane 1 : rat brain whole tissue lysates
      Lane 2 : rat spinal cord whole tissue lysates
      Lane 3 : mouse brain whole tissue lysates
      Lane 4 : mouse spinal cord whole tissue lysates
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-68kDa Neurofilament/NF-L antibody [DA2] (ab7255)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-68kDa Neurofilament/NF-L antibody [DA2] (ab7255)
      IHC image of ab7255 staining in human hippocampus formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab7255, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    • Immunocytochemistry/ Immunofluorescence - Anti-68kDa Neurofilament/NF-L antibody [DA2] (ab7255)
      Immunocytochemistry/ Immunofluorescence - Anti-68kDa Neurofilament/NF-L antibody [DA2] (ab7255)

      Ab7255 staining 68kDa Neurofilament/NF-L in rat frontal cortex sections by Immunocytochemistry/Immunofluorescence.  Samples were incubated with primary antibody at 1/5000 dilution (red) and chicken polyclonal to GFAP. 

    • Western blot - Anti-68kDa Neurofilament/NF-L antibody [DA2] (ab7255)
      Western blot - Anti-68kDa Neurofilament/NF-L antibody [DA2] (ab7255)
      Lane 1 : Coomassie Blue stain
      Lane 2 : Heavy neurofilament antibody
      Lane 3 : Anti-160 kD Neurofilament Medium antibody [3H11] (ab7256)
      Lane 4 : Anti-68kDa Neurofilament/NF-L antibody [DA2] (ab7255)
      Lane 5 : alpha Internexin antibody
      Lane 6 : GFAP antibody

      All lanes : Cytoskeletal homogenates of rat spinal cord
    • Flow Cytometry - Anti-68kDa Neurofilament/NF-L antibody [DA2] (ab7255)
      Flow Cytometry - Anti-68kDa Neurofilament/NF-L antibody [DA2] (ab7255)
      Overlay histogram showing SH-SY5Y cells stained with ab7255 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab7255, 1/100 dilution) 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. This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

    参考文献 (5)

    ab7255 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab7255 は 5 報の論文で使用されています。

    • Rupprecht A  et al. Uncoupling protein 2 and 4 expression pattern during stem cell differentiation provides new insight into their putative function. PLoS One 9:e88474 (2014). WB ; Mouse . PubMed: 24523901
    • Roher AE  et al. Subjects harboring presenilin familial Alzheimer's disease mutations exhibit diverse white matter biochemistry alterations. Am J Neurodegener Dis 2:187-207 (2013). PubMed: 24093083
    • Dorval V  et al. Gene network and pathway analysis of mice with conditional ablation of Dicer in post-mitotic neurons. PLoS One 7:e44060 (2012). PubMed: 22952873
    • Zhang L  et al. {gamma}-Diketone Axonopathy: Analyses of Cytoskeletal Motors and Highways in CNS Myelinated Axons. Toxicol Sci 117:180-9 (2010). WB ; Rat . PubMed: 20554699
    • Edson KJ  et al. Identification of a tektin-like protein associated with neurofilaments in the developing chick nervous system. J Neurosci Res 30:105-15 (1991). PubMed: 1795395

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    Question

    What were the Western blot conditions used for the blot on the datasheet?

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    Answer

    Thank you for your enquiry. The membrane used was 0.45micron nitrocellulose, blocked in 5% non fat milk in TBS, antibody incubations in TBS +0.1% tween 20, ab7255 conc was equivalent of 1:500 from our standard tissue culture supernatant prep and incubation time was 1hr at RT with shaking. Detection in this case was with Goat anti-mouse alkaline phosphatase conjugate and development was with BCIP/NBT. The blot was made quite a while ago, probably about 1993, and if we were doing it today we would use chemiluminesnce and probably 1:1,000 to 1:2,000 ab concentration. I hope this information helps. Please do not hesitate to contact us again if you need anything further.

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    Question

    Hi, I'm interested in finding a good NF-L antibody. I am curious about the amount you reccomend using for immunoblotting (i.e. what kind of dilution, 1:1000, 1:200, 1:5000, etc. etc.) Normally the data sheet indicates this information, but I did not see it there. Thanks.

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    Answer

    Dear Sir/Madam: Our anti-NF-L monoclonal antibody (catalog# NB 300-132) isunpurified mouse ascites (50 ul). We suugest a working dilution of 1:1,000 for ECL. Best Regards, Kate Lynott

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