Overview

  • Product name
    Anti-NG2 antibody [LHM 2]
    See all NG2 primary antibodies
  • Description
    Mouse monoclonal [LHM 2] to NG2
  • Host species
    Mouse
  • Tested applications
    Suitable for: IHC-FoFr, WB, IP, IHC-Fr, Flow Cytmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Tissue/ cell preparation (Human - a 375P cell crude extract).

  • General notes
    The LHM2 antibody is a very good marker for melanoma (>90% stained). LHM2 also stains basal cell carcinomas and subpopulation of basal keratinocytes. LHM2 can be used as a diagnostic panel for immunopathology of malignant melanoma and potential candidates for immunoimaging and immunotherapy of disseminated melanoma. The originators of this antibody report that the MCSP is a cell surface marker specific for epidermal stem cells (J. Invest. Dermatol. in press). Potential applications of the LHM2 antibody include use for separation/isolation of epidermal stem cells, detection of stem cells in skin biopsies or epithelial cell cultures and optimisation of culture conditions to retain epidermal stem cell population in in vitro cultures.

Properties

Applications

Our Abpromise guarantee covers the use of ab20156 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 Use at an assay dependent concentration. PubMed: 21970342
WB Use at an assay dependent concentration. Predicted molecular weight: 240 kDa.
IP Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

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

References

This product has been referenced in:
  • Kim JM  et al. Perivascular Progenitor Cells Derived From Human Embryonic Stem Cells Exhibit Functional Characteristics of Pericytes and Improve the Retinal Vasculature in a Rodent Model of Diabetic Retinopathy. Stem Cells Transl Med 5:1268-76 (2016). Human . Read more (PubMed: 27388242) »
  • Loibl M  et al. Direct cell-cell contact between mesenchymal stem cells and endothelial progenitor cells induces a pericyte-like phenotype in vitro. Biomed Res Int 2014:395781 (2014). ICC/IF . Read more (PubMed: 24563864) »
See all 12 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Vein Vessel Wall)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer, pH 9
Permeabilization
Yes - Dako Diluent and Blocking buffer
Specification
Vein Vessel Wall
Fixative
Formaldehyde

Dr. Piotr Kupczyk

Verified customer

Submitted Aug 08 2018

Answer

Thank you for your inquiry.

Indeed, you are right. The three antibodies, ab78284, ab83508 and ab20156, are the only antibodies against NG2 which have been tested in flow cytometry and on human cells. Of those three, the ab20156 is the only one provided in PBS only.

Please do let me know however for what application you need the antibody exactly and I might be able to advise you further or to provide you a testing discount.

I can also recommend you our antibody purification kits if you are interested in purifying an antibody by yourself - https://www.abcam.com/index.html?pageconfig=resource&rid=13489

I hope this information is helpful. Please do not hesitate to contact us again if you have any further question.

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Answer

Thank you for your email and for your telephone enquiry earlier today.

As discussed on the telephone, PDGF receptor beta is a membrane protein, and so I would recommend that detection with ab35270 would not usuallyrequire permeablization. Though this may require some individual optimization.

NG2 is an apical cell membrane protein. Reviewing the details after our phone conversation,as this is a membrane protein, it will not require permeabilization. However, as this protein can alsorelocate to other parts of the cell, such as thelamellipodia, I can recommend you may require a gentle permeabilization step. A strong detergent like Triton may not be suitable, as this can disrupt membrane proteins and significantly affect the staining. I can suggest it would be more appropriate to include more gentle detergent such as 0.2% Tween or saponin in the antibody dilution buffers and wash buffers. This will help to keep the cells permeabilised. Again, this is a guideline only and may require some individual optimization.

Usually for membrane proteinswe would not recommend to use organic solventsto fix the samples, as this also permeabilizes the cells and can cause disruption to antibody epitopes on membrane proteins.However, I do notice that an Abreview on teh datasheet forthe NG2 antibody indicates that acetone fixation worked best in frozen sections. This does highlight and indicate that although we canprovide recommendations, individual optimization is sometimes required.

https://www.abcam.com/index.html?datasheet=20156&tab=abreviews&intabreviewid=6408

I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

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Question
Answer

DISCOUNT CODE for AB85308: Expiration date: I am very pleased to hear you would like to accept our offer and test ab20156 and/or ab85308 in IHC on rabbit. This code will give you 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for IHC in rabbit and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (skin)
Specification
skin
Blocking step
BSA as blocking agent for 15 minute(s) · Concentration: 3%

Dr. Sonja Muffler

Verified customer

Submitted Dec 14 2006

Answer

The antibody was tested in IHC on keratinocytes in the epidermis in the paper Ghali L et al. Epidermal and hair follicle progenitor cells express melanoma-associated chondroitin sulfate proteoglycan core protein. J Invest Dermatol 122:433-42 (2004). PubMed: 15009727 I'm afraid I do not have access to this paper to find out the protocol used to test the antibody. I suggest you try a range of dilutions and incubation times (1:50,1:100, 1:200, 1:500, 1:1000 for example). The following references have also used this clone: Kupsch JM et al. Isolation of human tumor-specific antibodies by selection of an antibody phage library on melanoma cells. Clin Cancer Res 5:925-31 (1999). PubMed: 10213230 Kupsch JM et al. Generation and selection of monoclonal antibodies, single-chain Fv and antibody fusion phage specific for human melanoma-associated antigens. Melanoma Res 5:403-11 (1995). PubMed: 8589614 I hope these references will help,

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