Overview

  • Product name
  • Description
    Rabbit polyclonal to NG2
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Horse, Cow, Dog, Macaque monkey, Chinese hamster, Orangutan
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 300 - 400 of Mouse NG2.

  • Positive control
    • This antibody gave a positive signal in the following tissue lysates: Mouse Brain; Mouse Hippocampus; Mouse Cerebellum; Mouse Spinal Cord; Rat Brain; Rat Spinal Cord. This antibody gave a positive result when used in the following methanol fixed cell lines: MALME-3M

Properties

Applications

Our Abpromise guarantee covers the use of ab129051 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-P Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 252 kDa (predicted molecular weight: 252 kDa). Abcam recommends using milk as the blocking agent - 3%
ICC/IF Use a concentration of 5 µg/ml.

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-P image of NG2 staining of human melanoma cells in mouse tissue sections using ab129051 (1: 500). The sections were deparaffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were then blocked using 1% BSA for 10 mins at 21°C. ab129051 was dikuted 1:500 using TBS containing BSA and Azide, incubated on the sections for 2 hours at 21°C. The secondary antibody used was Goat polyclonal to Rabbit IgG conjugated to Biotin (1:250).

    See Abreview

  • ICC/IF image of ab129051 stained Malme-3M cells. The cells were 100% methanol fixed (5 min) then permeabilised using 0.1% PBS-Triton and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to further permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab129051 at 5µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) 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-NG2 antibody (ab129051) at 1 µg/ml (Milk blocking - 3%)

    Lane 1 : Brain (Mouse) Tissue Lysate
    Lane 2 : Mouse Hippocampus Tissue Lysate
    Lane 3 : Cerebellum Mouse Tissue Lysate
    Lane 4 : Spinal Cord (Mouse) Tissue Lysate
    Lane 5 : Brain (Rat) Tissue Lysate
    Lane 6 : Spinal Cord (Rat) Tissue Lysate

    Lysates/proteins at 25 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 252 kDa
    Observed band size: 252 kDa


    Exposure time: 20 minutes


    Abcam recommends using milk as the blocking agent - 3%. This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab129051 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

References

This product has been referenced in:
  • Beckmann N  et al. Brain region-specific enhancement of remyelination and prevention of demyelination by the CSF1R kinase inhibitor BLZ945. Acta Neuropathol Commun 6:9 (2018). Read more (PubMed: 29448957) »
  • Gonçalves AF  et al. Evidence of renal angiomyolipoma neoplastic stem cells arising from renal epithelial cells. Nat Commun 8:1466 (2017). Read more (PubMed: 29133867) »

See all 2 Publications for this product

Customer reviews and Q&As

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry free floating
Sample
Mouse Tissue sections (Brain)
Specification
Brain
Username

Yacine Tensaouti

Verified customer

Submitted Jun 21 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Pig Cell (bone marrow-mesenchymal stem cell)
Permeabilization
Yes - saponin 0,1%
Specification
bone marrow-mesenchymal stem cell
Blocking step
Protein block serum-free ready to use (DAKO) + saponin 0,1% as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0% · Temperature: 25°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Aug 23 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 3% · Temperature: RT°C
Sample
Rat Tissue sections (brain)
Specification
brain
Permeabilization
Yes - tween 20 0.1%
Fixative
Formaldehyde
Username

Mr. Samuel Valable

Verified customer

Submitted Aug 30 2013

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Sample
Mouse Tissue sections (Brain)
Specification
Brain
Permeabilization
No
Fixative
Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

Submitted Jun 07 2013

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

Mr. Carl Hobbs

Verified customer

Submitted Jun 07 2013

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 (Human melanoma cells in mouse)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Human melanoma cells in mouse
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Fixative
Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

Submitted May 02 2013

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

Sign up