Recombinant
RabMAb

Recombinant Anti-68kDa Neurofilament/NF-L antibody [EPR22035-112] (ab223343)

Overview

  • Product name

    Anti-68kDa Neurofilament/NF-L antibody [EPR22035-112]
    See all 68kDa Neurofilament/NF-L primary antibodies
  • Description

    Rabbit monoclonal [EPR22035-112] to 68kDa Neurofilament/NF-L
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, IHC-Fr, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Mouse 68kDa Neurofilament/NF-L aa 350 to the C-terminus. The exact sequence is proprietary.
    Database link: P08551

  • Positive control

    • WB: Human, mouse and rat brain lysate. IHC-P: Human, mouse and rat cerebellum tissue. IHC-Fr: Mouse and rat cerebellum tissue. IP: Mouse brain lysate.
  • General notes

    Previously labelled as 68kDa Neurofilament. 

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR22035-112
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab223343 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
WB 1/1000. Detects a band of approximately 68 kDa (predicted molecular weight: 62 kDa).
IHC-P 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr 1/500.
IP 1/30.

Target

  • Function

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

    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.
  • Sequence similarities

    Belongs to the intermediate filament family.
  • Domain

    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.
  • Post-translational
    modifications

    O-glycosylated.
    Phosphorylated in the Head and Rod regions by the PKC kinase PKN1, leading to inhibit polymerization.
  • Information by UniProt
  • Database links

  • Alternative names

    • 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

Images

  • Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen sectioned mouse cerebellum tissue labeling 68kDa Neurofilament/NF-L with ab223343 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution (green). Positive staining in mouse cerebellum (PMID: 17598884) is observed. The nuclear counter stain is DAPI (blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

    Heat mediated antigen retrieval using sodium citrate buffer (10 mM citrate pH 6.0 + 0.05% Tween-20).

  • Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen sectioned rat cerebellum tissue labeling 68kDa Neurofilament/NF-L with ab223343 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution (green). Positive staining in rat cerebellum is observed. The nuclear counter stain is DAPI (blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

    Heat mediated antigen retrieval using sodium citrate buffer (10 mM citrate pH 6.0 + 0.05% Tween-20).

  • All lanes : Anti-68kDa Neurofilament/NF-L antibody [EPR22035-112] (ab223343) at 1/1000 dilution

    Lane 1 : Human brain lysate
    Lane 2 : Mouse brain lysate
    Lane 3 : Rat brain lysate
    Lane 4 : Mouse liver lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 62 kDa
    Observed band size: 68 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking and dilution buffer: 5% NFDM/TBST.

    Negative control: Mouse liver (PMID: 2120242).

  • Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue labeling 68kDa Neurofilament/NF-L with ab223343 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining of nerve fibers in rat cerebellum (PMID: 28190929) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

    Heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue labeling 68kDa Neurofilament/NF-L with ab223343 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining of nerve fibers in mouse cerebellum (PMID: 28190929) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

    Heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded human cerebellum tissue labeling 68kDa Neurofilament/NF-L with ab223343 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining of nerve fibers in human cerebellum (PMID: 28190929) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

    Heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • 68kDa Neurofilament/NF-L was immunoprecipitated from 0.35 mg of mouse brain lysate with ab223343 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab223343 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

    Lane 1: Mouse brain lysate 10 μg (Input).
    Lane 2: ab223343 IP in mouse brain lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab223343 in mouse brain lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 10 seconds.

  • Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen sectioned rat liver tissue labeling 68kDa Neurofilament/NF-L with ab223343 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution. No staining in rat liver is observed. The nuclear counter stain is DAPI (blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

    Heat mediated antigen retrieval using sodium citrate buffer (10 mM citrate pH 6.0 + 0.05% Tween-20).

    Negative control :Rat liver (PMID: 2120242).

  • Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen sectioned mouse liver tissue labeling 68kDa Neurofilament/NF-L with ab223343 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution. No staining observed in mouse liver. The nuclear counter stain is DAPI (blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

    Heat mediated antigen retrieval using sodium citrate buffer (10 mM citrate pH 6.0 + 0.05% Tween-20).

    Negative control: mouse liver (PMID: 2120242).

References

ab223343 has not yet been referenced specifically in any publications.

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