Astana Biomed Group, an authorized Abcam distributor in Central Asia

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

Key features and details

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

Overview

  • Product name

    Anti-68kDa Neurofilament/NF-L antibody [DA2]
    See all 68kDa Neurofilament/NF-L primary antibodies

  • Description

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

  • Host species

    Mouse

  • Specificity

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

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Rat
    IHC-P
    Human
    WB
    Rat
    See all applications and species data

  • Immunogen

    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.

  • Epitope

    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.

  • General notes

    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. 

    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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • 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.

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