Overview

  • Product name

  • Description

    Rabbit polyclonal to NG2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-FoFr, ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human NG2 aa 1300-1400 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab96112)

  • Positive control

    • This antibody gave a positive signal in Human Skin Tissue Lysate - adult normal tissue

Properties

Applications

Our Abpromise guarantee covers the use of ab83178 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 1/100.
ICC/IF 1/200.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 300 kDa (predicted molecular weight: 251 kDa).

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

  • IHC-FoFr image of NG2 staining on sections of rat cortex post injury using ab83178 (1:100). The brain was perfusion fixed using 4% PFA and the sections were permeabilized using 0.1% TritonX in 0.1% PBS. The sections were then blocked using 10% donkey serum for 1hour at 24°C. ab83178 was diluted 1:100 and incubated with the sections for 24 hours at 4°C. The secondary antibody used was donkey polyclonal to rabbit IgG conjugated to Alexa Fluor 488 (1:1000).

  • ICC/IF image of ICC/IF staining on miced glia culture cells using ab83178 (1:200). The mixed glia culture was fixed with PFA and the cells were then blocked with 10% donkey serum for 1 hour at 24°C. The cells were then incubated with ab83178 was diluted by 1:200 using 0.3% TritonX with 0.1% PBS and 10% donkey serum. The cells were then incubated with donkey polyclonal to antirabbit conjuaged to Alexa 488 (1:1000). DAPI was used to stain the nucleus.

    See Abreview

  • Anti-NG2 antibody (ab83178) at 1 µg/ml + Human skin tissue lysate - total protein (ab30166) at 10 µg

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

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 251 kDa
    Observed band size: 300 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 190 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 3 minutes


    Human Chondroitin sulfate proteoglycan 4 precursor (NG2) contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.

References

This product has been referenced in:

  • Zeng J  et al. TRIM9-Mediated Resolution of Neuroinflammation Confers Neuroprotection upon Ischemic Stroke in Mice. Cell Rep 27:549-560.e6 (2019). Read more (PubMed: 30970257) »
  • Giacci MK  et al. Oligodendroglia Are Particularly Vulnerable to Oxidative Damage after Neurotrauma In Vivo. J Neurosci 38:6491-6504 (2018). Read more (PubMed: 29915135) »
See all 6 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Pig Cell (brain Pericytes)
Permeabilization
Yes - 0,1% TritonX
Specification
brain Pericytes
Blocking step
Serum as blocking agent for 45 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative
Paraformaldehyde

Mrs. Verena Ledwig

Verified customer

Submitted Mar 09 2016

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Sample
Rat Tissue sections (Rat Brain post injury)
Specification
Rat Brain post injury
Permeabilization
Yes - 0.1% TritonX in 0.1% PBS
Fixative
Paraformaldehyde

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted Sep 27 2013

Answer

Merci de nous avoir contactés. Je suis désolé d'apprendre que vous avez des difficultés à obtenir de bons résultats avec ab83178.

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 6 mois, 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:

- cet anticorps a été caractérisé en utilisant comme agent bloquant 5% de BSA (1 heure à température ambiante). Je recommanderais d'essayer ces conditions, y compris pour la solution de dilution des 2 anticorps, primaire et secondaire.

- les conditions d'incubation recommandées sont sur la nuit à 4ºC.

- pour le transfert je recommanderais pour les protéines à haut poids moléculaire telle que la NG2 d'appliquer un faible voltage (25 V) sur la nuit ou 24 heures, suivi par un "boost" à 70 V pendant 1 heure.
Vous pouvez également diminuer ou éviter totalement l'emploi de méthanol qui a tendance à "laver" les protéines.
Enfin vous pouvez augmenter la quantité de SDS jusqu'à 0.05%.

J'espère que ces informations vous aideront. Si nos suggestions ne permettent pas d'améliorer vos résultats, n'hésitez pas à me recontacter pour plus d'assistance.

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Glial culture)
Specification
Glial culture
Fixative
Paraformaldehyde
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 Jul 09 2012

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