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Neuroscience Cell Type Marker Glia marker Oligodendrocyte marker

Anti-CNPase antibody [11-5B] (ab6319)

Price and availability

342 727 ₸

Availability

Order now and get it on Tuesday March 02, 2021

Anti-CNPase antibody [11-5B] (ab6319)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)

Key features and details

  • Mouse monoclonal [11-5B] to CNPase
  • Suitable for: Flow Cyt, ICC, IHC-P, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG1

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Overview

  • Product name

    Anti-CNPase antibody [11-5B]
    See all CNPase primary antibodies
  • Description

    Mouse monoclonal [11-5B] to CNPase
  • Host species

    Mouse
  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC
    Human
    IHC-P
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data
  • Immunogen

    Full length native protein (purified) corresponding to Human CNPase.

  • Positive control

    • WB: HeLa and Hap1 cell lysates; Human and Mouse Spinal Cord and Brain tissue lysates; Rat Brain tissue lysate. IHC-P: FFPE human cerebral cortex tissue sections. ICC: SKNSH cell line
  • General notes

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

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 or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 6.97% L-Arginine
  • Concentration information loading...
  • Purity

    IgG fraction
  • Clonality

    Monoclonal
  • Clone number

    11-5B
  • Isotype

    IgG1
  • Research areas

    • Neuroscience
    • Cell Type Marker
    • Glia marker
    • Oligodendrocyte marker
    • Tags & Cell Markers
    • Cell Type Markers
    • Neuroscience Markers
    • Glial
    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Other

Images

  • Western blot - Anti-CNPase antibody [11-5B] (ab6319)
    Western blot - Anti-CNPase antibody [11-5B] (ab6319)
    All lanes : Anti-CNPase antibody [11-5B] (ab6319) at 5 µg/ml

    Lane 1 : Wild-type HAP1 cell lysate at 40 µg
    Lane 2 : CNPase knockout HAP1 cell lysate at 40 µg
    Lane 3 : Human brain whole cell lysate at 20 µg

    Predicted band size: 48 kDa



    Lanes 1 - 3: Merged signal (red and green). Green - ab6319 observed at 48 kDa. Red - loading control, ab181602, observed at 37 kDa.

    ab6319 was shown to recognize CNPase in wild-type HAP1 cells as signal was lost at the expected MW in CNPase knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and CNPase knockout samples were subjected to SDS-PAGE. Ab6319 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 5 μg/ml 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CNPase antibody [11-5B] (ab6319)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CNPase antibody [11-5B] (ab6319) This image is courtesy of an abreview submitted by Carl Hobbs, King's College London, United Kingdom

    IHC-P image of CNPase staining on rat brain sections using ab6319 (1/1600). heat mediated antigen retrieval on paraffin embedded sections was performed using citric acid. The sections were then blocked with 1% BSA for 10 min at 21°C. The primary antibody was incubated for 16 hours at 21°C. The sections were then incubated in Goat anti-mouse (Biotin) at 1:200.

    See Abreview

  • Western blot - Anti-CNPase antibody [11-5B] (ab6319)
    Western blot - Anti-CNPase antibody [11-5B] (ab6319)
    All lanes : Anti-CNPase antibody [11-5B] (ab6319)

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : CNP knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 48 kDa
    Observed band size: 48 kDa



    Lanes 1- 2: Merged signal (red and green). Green - ab6319 observed at 48 kDa. Red - Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) observed at 37 kDa.

     ab6319 was shown to react with CNPase in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264949 (knockout cell lysate ab256877) was used. Wild-type HeLa and CNP knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab6319 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) overnight at 4°C at a 5 µg/ml and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye®800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye®680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-CNPase antibody [11-5B] (ab6319)
    Western blot - Anti-CNPase antibody [11-5B] (ab6319)
    All lanes : Anti-CNPase antibody [11-5B] (ab6319) at 1/100 dilution

    Lane 1 : Human spinal cord tissue lysate - total protein (ab29188)
    Lane 2 : Human brain tissue lysate - total protein (ab29466)
    Lane 3 : Spinal Cord (Mouse) Tissue Lysate
    Lane 4 : Brain (Mouse) Tissue Lysate
    Lane 5 : Spinal Cord (Rat) Tissue Lysate
    Lane 6 : Brain (Rat) Tissue Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 48 kDa


    Exposure time: 1 minute


    This antibody was raised against full length native CNPase and is predicted to recognize both isoforms. The predicted molecular weights of isoforms CNPI and CNPII are 45- and 48-kDa respectively.
  • Immunocytochemistry - Anti-CNPase antibody [11-5B] (ab6319)
    Immunocytochemistry - Anti-CNPase antibody [11-5B] (ab6319) This image is courtesy of an anonymous abreview.
    ab6319 staining CNPase in rat oligodendrocytes by Immunocytochemistry/ Immunofluorescence.
    Cells were fixed in paraformaldehyde, blocked using 5% serum for 10 minutes at 25°C, then incubated with ab6319 at a 1/200 dilution for 2 hours at 25°C. The secondary used was a goat anti-mouse Cy3 conjugated polyclonal at a 1/100 dilution.

    See Abreview

  • Flow Cytometry - Anti-CNPase antibody [11-5B] (ab6319)
    Flow Cytometry - Anti-CNPase antibody [11-5B] (ab6319)

    Overlay histogram showing SH-SY5Y cells stained with ab6319 (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 (ab6319, 2µg/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. This antibody gave a positive signal in SH-SY5Y cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CNPase antibody [11-5B] (ab6319)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CNPase antibody [11-5B] (ab6319)

    IHC image of CNPase staining in human cerebral cortex formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ 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 ab6319, 5µg/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 - Anti-CNPase antibody [11-5B] (ab6319)
    Immunocytochemistry - Anti-CNPase antibody [11-5B] (ab6319) This image is courtesy of an anonymous Abreview

    ab6319 staining CNPase in the rat oligodendrocytes by ICC/IF (Immunoytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with methanol and blocked with 5% BSA for 1 hour at 37°C. Samples were incubated with primary antibody (1/100 in PBS) for 18 hours at 4°C. An Alexa Fluor® 594-conjugated Goat anti-mouse IgG polyclonal (1:200) was used as the secondary antibody.

    See Abreview

  • Western blot - Anti-CNPase antibody [11-5B] (ab6319)
    Western blot - Anti-CNPase antibody [11-5B] (ab6319) This image is courtesy of an anonymous Abreview
    All lanes : Anti-CNPase antibody [11-5B] (ab6319) at 1/750 dilution

    All lanes : Spinal Cord homogenate (whole tissue lysate)

    Lysates/proteins at 2 µg per lane.

    Secondary
    All lanes : HRP conjugated sheep anti-mouse IgG

    Predicted band size: 48 kDa
    Observed band size: 45,47 kDa
    why is the actual band size different from the predicted?

    See Abreview

  • Immunocytochemistry - Anti-CNPase antibody [11-5B] (ab6319)
    Immunocytochemistry - Anti-CNPase antibody [11-5B] (ab6319)

    ab6319 stained in SKNSH cells. Cells were fixed with 100% methanol (10 min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% Triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab6319 at 10µg/ml and ab6046 (Rabbit polyclonal to beta Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CNPase antibody [11-5B] (ab6319)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CNPase antibody [11-5B] (ab6319) This image is courtesy of an abreview submitted by Carl Hobbs, King's College London, United Kingdom

    ab6319 staining CNPase in Dog Cerebellum tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/1500 in blocking buffer) for 2 hours at 21°C. A Biotin-conjugated Goat anti-mouset IgG polyclonal (1/200) was used as the secondary antibody.

    See Abreview

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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