Recombinant
RabMAb

Recombinant Anti-NG2 antibody [EPR20244] (ab183929)

Rabbit recombinant monoclonal NG2 antibody [EPR20244]. Validated in WB, IHC, ICC/IF and tested in Human. Immunogen corresponding to recombinant fragment.

Overview

  • Product name

    Anti-NG2 antibody [EPR20244]
    See all NG2 primary antibodies
  • Description

    Rabbit monoclonal [EPR20244] to NG2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human NG2 aa 150-450. The exact sequence is proprietary.
    Database link: Q6UVK1

  • Positive control

    • WB: A-375 and MeWo whole cell lysates; Human skin lysate. IHC-P: Human melanoma and glioma tissues. ICC/IF: A-375 and MeWo cells.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab183929 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 1/1000. Predicted molecular weight: 251 kDa.
ICC/IF 1/100.
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

  • Immunohistochemical analysis of paraffin-embedded human melanoma tissue labeling NG2 with ab183929 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Membranous and weak cytoplasmic staining on tumor cells of human melanoma (PMID: 24258997). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, Goat Anti-Rabbit IgG H&L (HRP) ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • All lanes : Anti-NG2 antibody [EPR20244] (ab183929) at 1/1000 dilution

    Lane 1 : A-375 (human malignant melanoma cell line), whole cell lysate
    Lane 2 : Human skin lysate
    Lane 3 : MeWo (human malignant melanoma cell line), whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Lanes 1 & 3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
    Lane 2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 251 kDa
    Observed band size: 270 kDa
    why is the actual band size different from the predicted?



     

    Exposure times: Lanes 1,3: 1 minute; Lane 2: 3 minutes.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    This image of lane 3 was performed on a BIO-RAD® ChemiDoc™ MP instrument.

     

     

  • Immunofluorescent analysis of 100% methanol-fixed A-375 (human malignant melanoma cell line) cells labeling NG2 with ab183929 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membrane staining on A-375 cell line.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human glioma tissue labeling NG2 with ab183929 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Membranous staining on blood vessels of human glioma (PMID: 24386429). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, Goat Anti-Rabbit IgG H&L (HRP) ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunofluorescent analysis of 100% methanol-fixed MeWo (human malignant melanoma cell line) cells labeling NG2 with ab183929 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membrane staining on MeWo cell line.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

References

ab183929 has not yet been referenced specifically in any publications.

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