Overview

  • Product name
    Anti-NG2 antibody [132.38]
    See all NG2 primary antibodies
  • Description
    Mouse monoclonal [132.38] to NG2
  • Host species
    Mouse
  • Tested applications
    Suitable for: WB, IP, ELISA, ICC/IF, IHC-Fr, IHC-FoFrmore details
  • Species reactivity
    Reacts with: Rat
    Does not react with: Mouse
  • Immunogen

    Cell extract of HEK293 cells expressing the D3 domain of rat NG2, corresponding to amino acids 1592-2222.

  • Positive control
    • This antibody gave a positive signal in the following rat tissue lysates: spinal cord; whole brain; hippocampus.
  • 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.

    The Protocols tab contains a Mouse on Mouse staining protocol with recommendations when using a mouse monoclonal antibody to stain mouse tissues and tips for reducing background.

Properties

Applications

Our Abpromise guarantee covers the use of ab50009 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 Use a concentration of 5 µg/ml. Detects a band of approximately 251 kDa (predicted molecular weight: 251 kDa).

Abcam recommends using BSA as the blocking agent.

IP Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
ICC/IF 1/200. PubMed: 18974869
IHC-Fr Use at an assay dependent concentration.
IHC-FoFr Use at an assay dependent concentration.

Target

  • Function
    Proteoglycan playing a role in cell proliferation and migration which stimulates endothelial cells motility during microvascular morphogenesis. May also inhibit neurite outgrowth and growth cone collapse during axon regeneration. Cell surface receptor for collagen alpha 2(VI) which may confer cells ability to migrate on that substrate. Binds through its extracellular N-terminus growth factors, extracellular matrix proteases modulating their activity. May regulate MPP16-dependent degradation and invasion of type I collagen participating in melanoma cells invasion properties. May modulate the plasminogen system by enhancing plasminogen activation and inhibiting angiostatin. Functions also as a signal transducing protein by binding through its cytoplasmic C-terminus scaffolding and signaling proteins. May promote retraction fiber formation and cell polarization through Rho GTPase activation. May stimulate alpha-4, beta-1 integrin-mediated adhesion and spreading by recruiting and activating a signaling cascade through CDC42, ACK1 and BCAR1. May activate FAK and ERK1/ERK2 signaling cascades.
  • Tissue specificity
    Detected only in malignant melanoma cells.
  • Sequence similarities
    Contains 15 CSPG (NG2) repeats.
    Contains 2 laminin G-like domains.
  • Post-translational
    modifications
    O-glycosylated; contains glycosaminoglycan chondroitin sulfate which are required for proper localization and function in stress fiber formation (By similarity). Involved in interaction with MMP16 and ITGA4.
    Phosphorylation by PRKCA regulates its subcellular location and function in cell motility.
  • Cellular localization
    Apical cell membrane. Cell projection > lamellipodium membrane. Localized at the apical plasma membrane it relocalizes to the lamellipodia of astrocytoma upon phosphorylation by PRKCA. Localizes to the retraction fibers. Localizes to the plasma membrane of oligodendrocytes.
  • Information by UniProt
  • Database links
  • Alternative names
    • 4732461B14Rik antibody
    • AN2 antibody
    • AN2 proteoglycan antibody
    • Chondroitin sulfate proteoglycan 4 (melanoma-associated) antibody
    • Chondroitin sulfate proteoglycan 4 antibody
    • Chondroitin sulfate proteoglycan NG2 antibody
    • Cspg4 antibody
    • Cspg4 chondroitin sulfate proteoglycan 4 antibody
    • CSPG4_HUMAN antibody
    • HMW-MAA antibody
    • HSN tumor-specific antigen antibody
    • Kiaa4232 antibody
    • MCSP antibody
    • MCSPG antibody
    • MEL-CSPG antibody
    • Melanoma chondroitin sulfate proteoglycan antibody
    • Melanoma-associated chondroitin sulfate proteoglycan antibody
    • MELCSPG antibody
    • MSK16 antibody
    • NG2 antibody
    see all

Images

  • ab76897 staining NG2 in rat brain tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue was fixed with paraformaldehyde, permeabilized with 0.3% Triton and blocked with 5% serum for 1 hour at 24°C. Samples were incubated with primary antibody (1/250) for 16 hours at 4°C. An Alexa Fluor®488 -conjugated Donkey anti-mouse IgG polyclonal (1/500) was used as the secondary antibody.

    See Abreview

  • ab50009 staining NG2 in an immunoreactive cell (rat hippocampal) by Immunohistochemistry (PFA fixed).
    Brains were postfixed (2–6 hours) in 4% PFA and placed in 30% sucrose (2 days) and sectioned (20–30 µm) with a cryostat. ab50009 was used at a 1/200 dilution. A goat anti-mouse-Alexa 594, 1/500, was used as the secondary antibody.
  • All lanes : Anti-NG2 antibody [132.38] (ab50009) at 1 µg/ml

    Lane 1 : Rat Brain Tissue Lysate
    Lane 2 : Rat Spinal Cord Tissue Lysate
    Lane 3 : Rat Hippocampus Tissue Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 251 kDa
    Observed band size: 270 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 31 kDa (possible non-specific binding), 41 kDa (possible non-specific binding)


    Exposure time: 20 minutes

References

This product has been referenced in:
See all 16 Publications for this product

Customer reviews and Q&As

1-10 of 12 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: sodium citrate pH 6.0
Permeabilization
Yes - 1% Triton-X
Specification
brain
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Fixative
Formalin

Abcam user community

Verified customer

Submitted Jan 24 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry
Sample
Mouse Cultured Cells (primary cells)
Permeabilization
Yes - Triton 0.2%
Specification
primary cells
Blocking step
NGS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 20°C
Fixative
acetone:methanol 1:1 with and without permeabilization and paraformaldehyde 4%

Ms. Renee Raaijmakers

Verified customer

Submitted Nov 28 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (brain)
Antigen retrieval step
None
Permeabilization
Yes - PBSTx 3x5mn RT
Specification
brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
Fixative
Paraformaldehyde

Yacine Tensaouti

Verified customer

Submitted Aug 02 2017

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Antigen retrieval step
None
Sample
Rat Tissue sections (brain)
Specification
brain
Permeabilization
Yes - 0.3% Triton-X
Fixative
Paraformaldehyde

Mrs. Olga Cravetchi

Verified customer

Submitted Mar 02 2015

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Brain)
Antigen retrieval step
None
Permeabilization
Yes - 0.1% Triton X-100
Specification
Brain
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Aug 26 2013

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate
Permeabilization
No
Specification
Brain
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted May 23 2013

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain)
Permeabilization
No
Specification
Brain
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted May 23 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Rat Tissue (Brain section)
Specification
Brain section
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% TritonX in 0.1% PBS
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted Feb 18 2013

Answer

Merci de ces précisions.

Eventuellement une autre fixation pourrait vous donner des meilleurs résultats, car la fixation acétone/méthanol fonctionne par précipitation et non pas crosslinking comme la PFA. Comme par contre nous avons des données que cet anticorps devrait fonctionner sur des coupes fixées PFA, je suis d'accord de vous remplacer cet anticorps avec un autre anticoprs en attendant que nous avions résolu si oui ou non cet anticorps fonctionne sur des coupes fixées PFA.

Commes autres anticorps contre le NG2, qui ne sont pas produits chez le lapin, nous avons les anticorps suivants:
- polyclonal de chèvre, ab101679, testé sur coupes de souris congélées et coupes paraffines
Click here (or use the following: https://www.abcam.com/index.html?datasheet=101679).
- monoclonal de souris, ab83508, testé sur la souris et en immunohistochimie
Click here (or use the following: https://www.abcam.com/index.html?datasheet=83508).

Veuillez me confirmer si vous accéptez le remplacment du produit ab50009 avec un unité gratuite de soit ab101679 ou de ab83508. N'hésitez pas non plus à nous contacter si vous avez des autres questions. Je me réjouis de votre réponse.

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Answer

Merci d'avoir pris le temps de remplir notre questionnaire et de nous avoir contactés. Je suis désolée d'apprendre que vous avez eu des difficultés à obtenir de bons résultats avec ab50009.
Nous avons eu un autre retour négative dernièrement sur cet anticorps et nous allons investiguer si nous trouvons l'origine de ce problème. Nous avons pas des autres indications qu'il y aurais un problème avec cet anticorps. Je peux vous assurer que la qualité de nos produits est très importante et que nous investiguons tous les problèmes.
Notre politique est d'offrir tout d'abord la meilleure assistance technique. Si nous concluons que le produit que vous avez reçu ne fonctionne pas comme mentionné sur sa fiche technique et que ce produit a été commandé sous 180 jours, nous nous ferons un plaisir de vous offrir un remplacement gratuit ou un avoir en compensation.

J'aimerais donc vous offrir quelques suggestions afin d'optimiser vos résultats. J'apprécierais si vous pouviez confirmer quelques détails de la procédure.

1.) Est-ce que vous avez testé aussi des autres méthodes de fixation comme l'acétone ou le méthanol tout seul?
2.) Pouvez vous confirmer que l'anticorps secondaire fonctionne avec des autres anticorps primaires de souris?
3.) Est-ce que vous pouvez nous confirmer la date de commande ou le numéro de commande de ce produit?
Si nous arrivons pas à résoudre ce problème, nous allons vous remplacer cet anticorps. Nous avons encore des autres anticorps de souris contre le NG2. Je me réjouis de votre réponse.

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1-10 of 12 Abreviews or Q&A

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