Anti-CNPase antibody [mAbcam 44289] (ab44289)

Overview

  • Product nameAnti-CNPase antibody [mAbcam 44289]See all CNPase primary antibodies ...
  • Description
    Mouse monoclonal [mAbcam 44289] to CNPase
  • Tested applicationsIP, Flow Cyt, IHC-FoFr, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Dog, Human
    Predicted to work with: Orangutan
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 350 to the C-terminus of Human CNPase.

    (Peptide available as ab19424.)

  • Positive control
    • This antibody gave a positive signal in the following Tissue Lysates: Human Brain, Mouse Brain, Mouse Spinal Cord, Rat Brain, Rat Spinal Cord
  • General notes

    We can conjugate this antibody to FITC for you (please see ab150257 for details).

Properties

Applications

Our Abpromise guarantee covers the use of ab44289 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
IP Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.
IHC-FoFr 1/300.
IHC-P 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 46,48 kDa (predicted molecular weight: 46,48 kDa).Can be blocked with Human CNPase peptide (ab19424).

Target

Anti-CNPase antibody [mAbcam 44289] images

  • Anti-CNPase antibody [mAbcam 44289] (ab44289) at 5 µg/ml + Brain (Human) Tissue Lysate - adult normal tissue (ab29466) at 10 µg

    Secondary
    Rabbit polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 46,48 kDa
    Observed band size : 46,48 kDa
  • IHC-P image of CNPase staining on human brain section using ab44289 (1:100). The paraffin embedded section was subjected to heat mediated antigen retrieval using citric acid. The sections were then blocked using 1% BSA at 21°C for 10 min. ab44289 was incubated at 21°C for 16 hours. The secondary antibody was goat polyclonal anti-mouse IgG conjugated to biotin (1:200)

    See Abreview

  • All lanes : Anti-CNPase antibody [mAbcam 44289] (ab44289) at 1 µg/ml

    Lane 1 : Brain (Mouse) Tissue Lysate
    Lane 2 : Spinal Cord (Mouse) Tissue Lysate
    Lane 3 : Brain (Rat) Tissue Lysate
    Lane 4 : Spinal Cord (Rat) Tissue Lysate
    Lane 5 : Brain (Human) Tissue Lysate - adult normal tissue (ab29466)

    Lysates/proteins at 10 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 : 46,48 kDa
    Observed band size : 46,48 kDa
    Additional bands at : 42 kDa,67 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 3 minutes
  • Overlay histogram showing SHSY-5Y cells stained with ab44289 (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 (ab44289, 1µg/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 SHSY-5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • CNPase was immunoprecipitated using 0.5mg Rat Brain whole tissue lysate, 5µg of Mouse monoclonal to CNPase and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Rat Brain whole tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab44289.
    Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
    Band: 48,46kDa: CNPase.
  • IHC-FoFr image of CNPase staining on rat dentate gyrus sections using ab44289(1:300). The sections used came from animals perfused fixed with Paraformaldehyde 4% with 15% of a solution of saturated picric acid, in phosphate buffer 0.1M. Following postfixation in the same fixative overnight, the brains were cryoprotected in sucrose 30% overnight. Brains were then cut using a cryostat and the immunostainings were performed using the ‘free floating’ technique.

    See Abreview

References for Anti-CNPase antibody [mAbcam 44289] (ab44289)

ab44289 has not yet been referenced specifically in any publications.

Product Wall

Merci de nous avoir contactés.

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Merci pour ce complément d'information qui nous sera très utile pour vérifier la spécificité de ces 2 anticorps.

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Je suis désolé d'apprendre que vous avez eu des difficultés à obtenir de bons résultats avec ab4674 et ab44289.
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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Rat Tissue sections (Brain)
Specification Brain
Fixative Paraformaldehyde
Antigen retrieval step None
Permeabilization No
Username

Dr. Sophie Pezet

Verified customer

Submitted Jul 04 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Dog Tissue sections (Cerebellum)
Specification Cerebellum
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 19 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human 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 14 2012

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Rat 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 12 2012

Thank you for providing the completed questionnaire form. I will be happy to help you resolve any issues you are having with ab44289. I just have a few comments: 1. It seems for storage you have diluted ab44289 50x in PBS before freezing?...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (brain)
Specification brain
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Dako Target Retreival Solution
Permeabilization No
Blocking step 5 minutes of peroxidase block then 10 minutes of protein block. These are ready-to-use reagents purchased from Dako as blocking agent for 45 minute(s) · Concentration: 100% · Temperature: 20°C
Username

Mr. Antibody Solutions

Verified customer

Submitted May 26 2010

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