Overview

  • Product nameAnti-CNPase antibody [11-5B]
    See all CNPase primary antibodies
  • Description
    Mouse monoclonal [11-5B] to CNPase
  • SpecificityReacts specifically with CNPase. In an immunoblotting assay, the antibody localizes both CNP1 (46kD) and CNP2 (48kD) bands. Immunohistochemical staining of paraffin, cryostat or vibratome sections of rat brain reveals selective staining of oligodendrocytes in the grey and white matter. Nerve cells and axons are not stained and astroglial cells do not appear to be labelled.
  • Tested applicationsIHC-FoFr, Flow Cyt, Conjugation, Dot Blot, ELISA, ICC, IHC-P, IHC-Fr, IP, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Dog, Human, Rhesus monkey
    Predicted to work with: Sheep, Rabbit, Cow, Pig

    Does not react with

    Chicken, Guinea pig
  • Immunogen

    Full length native protein (purified) (Human).

  • Positive control
    • This antibody gave a positive signal in Western Blot, when tested against Human, Mouse and Rat Spinal Cord and Brain tissue lysates. This antibody gave a positive result when used in the following formaldehyde fixed cell lines: SKNSH

Properties

Applications

Our Abpromise guarantee covers the use of ab6319 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
IHC-FoFr 1/250. PubMed: 19259393
Flow Cyt Use 2µg for 106 cells.
Conjugation Use at an assay dependent dilution.
Dot Blot Use at an assay dependent dilution.
ELISA Use at an assay dependent dilution.
ICC Use at an assay dependent dilution.
IHC-P 1/2000 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IHC-Fr Use at an assay dependent dilution.
IP Use at an assay dependent dilution.
WB Use a concentration of 5 µg/ml. Detects a band of approximately 48 kDa (predicted molecular weight: 48 kDa).
ICC/IF 1/200. PubMed: 17464316

Target

Anti-CNPase antibody [11-5B] images

  • ICC/IF image of ab6319 stained SKNSH cells. The cells were 4% formaldehyde fixed (10 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 ab6319 at 10µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- mouse (ab150117) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

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

    Lane 1 : Spinal Cord (Human) Tissue Lysate - adult normal tissue (ab29188)
    Lane 2 : Brain (Human) Tissue Lysate - adult normal tissue (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
    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.
  • 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

  • ab6319 at a 1/2000 dilution staining mouse brain tissue sections by Immunohistochemistry (Frozen sections). The tissue was paraformaldehyde fixed and blocked with 2% BSA. The antibody was incubated with the tissue for 10 hours and then bound antibody was detected using a biotinylated goat anti-mouse antibody.

    CNPase staining in the striatum is shown in the image.

    This image is courtesy of an Abreview submitted by Bing Lang on 19 April 2006.

    See Abreview

  • ab6319 at a 1/200 dilution staining rat spinal cord cells by ICC/IF. The cells were ethanol fixed and blocked with 5% serum. The antibody was incubated with the cells for 1 hour and then bound antibody was detected using an Alexa Fluor® 488 conjugated Goat anti-mouse antibody.

    This image is courtesy of an Abreview submitted by Nancy Nutile-McMenemy.

    See Abreview

  • ab6319 at a 1/200 dilution staining rat spinal cord tissue sections from a 4% PFA transcardially perfused animal by Immunohistochemistry (Frozen sections). The tissue was paraformaldehyde fixed and incubated with the antibody for 18 hours. Bound antibody was detected using an HRP conjugated goat anti-mouse polyclonal antibody.

    This image is courtesy of an Abreview submitted by Nancy Nutile-McMenemy.

    See Abreview

  • All lanes : Anti-CNPase antibody [11-5B] (ab6319) at 1/500 dilution

    Lane 1 : 40ug rat brain homegenate (whole tissue lysate) animal #661
    Lane 2 : 40ug rat brain homegenate (whole tissue lysate) animal #655
    Lane 3 : 40ug rat brain homegenate (whole tissue lysate) animal #668
    Lane 4 : 40ug rat brain homegenate (whole tissue lysate) animal #662
    Lane 5 : 40ug rat brain homegenate (whole tissue lysate) animal #659
    Lane 6 : 40ug rat brain homegenate (whole tissue lysate) animal #670

    Secondary
    HRP conjugated goat anti-mouse antibody
    developed using the ECL technique

    Performed under reducing conditions.

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


    Exposure time : 5 minutes

    This image is courtesy of an Abreview submitted by Ms Nancy Nutile-McMenemy

    See Abreview

  • All lanes : Anti-CNPase antibody [11-5B] (ab6319) at 1/750 dilution

    Lane 1 : Spinal Cord homogenate (whole tissue lysate)
    Lane 2 : Spinal Cord homogenate (whole tissue lysate)
    Lane 3 : Spinal Cord homogenate (whole tissue lysate)

    Lysates/proteins at 2 µg per lane.

    Secondary
    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?)

    This image is courtesy of an anonymous Abreview

    See Abreview

  • ab6319 staining mouse brain tissue sections by IHC-FoFr.  Sections were PFA fixed and permeabilized in 0.1% Triton X-100 prior to blocking in 0.5% TNB for 30 minutes at 25°C. The primary antibody was diluted 1/250 and incubated with the sample for 18 hours at 25°C.  An Alexa Fluor® 488 conjugated goat anti-mouse antibody, diluted 1/250, was used as the secondary.

    Image demonstrates a 2-D depth projection through the superficial cortex.

    See 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

  • 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.

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

This product has been referenced in:
  • Marion-Poll L  et al. Fluorescence-activated sorting of fixed nuclei: a general method for studying nuclei from specific cell populations that preserves post-translational modifications. Eur J Neurosci 39:1234-44 (2014). Flow Cyt ; Mouse . Read more (PubMed: 24713002) »
  • Xu YH  et al. Multiple pathogenic proteins implicated in neuronopathic Gaucher disease mice. Hum Mol Genet N/A:N/A (2014). Mouse . Read more (PubMed: 24599400) »

See all 26 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Sample Mouse Tissue sections (Brain)
Specification Brain
Permeabilization Yes - 0.05% Tween20
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Sep 24 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 100 µg
Gel Running Conditions Reduced Denaturing (10% Gel)
Sample Rat Tissue lysate - whole (white matter)
Specification white matter
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Username

Abcam user community

Verified customer

Submitted Sep 16 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Sample Rat Cell (oligodendrocytes)
Specification oligodendrocytes
Permeabilization Yes - Methanol
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Sep 16 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Rhesus monkey Tissue sections (Brain)
Specification Brain
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH6.0
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 20% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Mar 11 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Rat Cell lysate - whole cell (Oligodendrocytes)
Loading amount 30 µg
Specification Oligodendrocytes
Gel Running Conditions Reduced Denaturing (12)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Feb 15 2013

DISCOUNT CODE: ***
Expiration date: March 14, 2013

I am very pleased to hear you would like to accept our offer and test ab6319 in samples of rhesus macaque tissue. To redeem this offer, please submit an Abreview for rhesus macaque and ...

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Thank you for your reply.

As your sections are so thin, you should be able to just add the PFA to the sections when they come out of the freezer and not have to worry about it coming to room temperature.

Please let me know if there is...

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Thank you for your reply.

Based on the images you sent, I think that there may be a fixation issue, as based on the CNPase image (intense staining in the upper right hand side), I think that the tissue may not be the tissue is not being fixed ...

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Thank you for providing that information.

These antibodies are covered under our Abpromise for six months and are guaranteed to work in IHC on a variety of samples. If we cannot resolve the issue you are having with the antibody then I would b...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Brain)
Specification Brain
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Username

Mr. Carl Hobbs

Verified customer

Submitted Jun 18 2012

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"