Overview

  • Product nameAnti-CD44v6 antibody [VFF-18]
    See all CD44v6 primary antibodies
  • Description
    Mouse monoclonal [VFF-18] to CD44v6
  • SpecificitySpecific for an epitope encoded by exon v6 on the variant portion of human CD44.
  • Tested applicationsSuitable for: WB, IHC-P, ICC, Flow Cyt, IHC-Frmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Fusion protein corresponding to Human CD44v6. Glutathione S Transferase (GST) fusion protein corresponding to the variable domain v6 of human CD44.
    Database link: P16070

  • EpitopeSpecific for an epitope encoded by exon v6 on the variant portion of human CD44.

Properties

Applications

Our Abpromise guarantee covers the use of ab78960 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 at an assay dependent concentration. Detects a band of approximately 91 kDa (predicted molecular weight: 82 kDa).
IHC-P Use a concentration of 1 - 2 µg/ml. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.
ICC Use at an assay dependent concentration.

Use on cell smears at an assay dependent dilution.

Flow Cyt Use 1µg for 106 cells.

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

IHC-Fr Use a concentration of 1 - 2 µg/ml.

Use at a concentration of 1 - 2 µg/ml. Use acetone fixation and the alkaline phosphatase-anti-alkaline-phosphatase (APAAP) or peroxidase anti-peroxidase (PAP) procedures.

Target

  • FunctionReceptor for hyaluronic acid (HA). Mediates cell-cell and cell-matrix interactions through its affinity for HA, and possibly also through its affinity for other ligands such as osteopontin, collagens, and matrix metalloproteinases (MMPs). Adhesion with HA plays an important role in cell migration, tumor growth and progression. In cancer cells, may play an important role in invadopodia formation. Also involved in lymphocyte activation, recirculation and homing, and in hematopoiesis. Altered expression or dysfunction causes numerous pathogenic phenotypes. Great protein heterogeneity due to numerous alternative splicing and post-translational modification events.
  • Tissue specificityIsoform 10 (epithelial isoform) is expressed by cells of epithelium and highly expressed by carcinomas. Expression is repressed in neuroblastoma cells.
  • Sequence similaritiesContains 1 Link domain.
  • DomainThe lectin-like LINK domain is responsible for hyaluronan binding.
  • Post-translational
    modifications
    Proteolytically cleaved in the extracellular matrix by specific proteinases (possibly MMPs) in several cell lines and tumors.
    N- and O-glycosylated. O-glycosylation contains more-or-less-sulfated chondroitin sulfate glycans, whose number may affect the accessibility of specific proteinases to their cleavage site(s). It is uncertain if O-glycosylation occurs on Thr-637 or Thr-638.
    Phosphorylated; activation of PKC results in the dephosphorylation of Ser-706 (constitutive phosphorylation site), and the phosphorylation of Ser-672.
  • Cellular localizationMembrane. Colocalizes with actin in membrane protrusions at wounding edges.
  • Information by UniProt
  • Database links
  • Alternative names
    • CD44 antibody
    • CD44 antigen antibody
    • CD44 molecule antibody
    • CD44_HUMAN antibody
    • CDw44 antibody
    • ECMR-III antibody
    • Epican antibody
    • Extracellular matrix receptor III antibody
    • GP90 lymphocyte homing/adhesion receptor antibody
    • Heparan sulfate proteoglycan antibody
    • Hermes antigen antibody
    • HUTCH I antibody
    • HUTCH-I antibody
    • Hyaluronate receptor antibody
    • MIC4 antibody
    • MUTCH I antibody
    • PGP 1 antibody
    • PGP-1 antibody
    • PGP-I antibody
    • Phagocytic glycoprotein 1 antibody
    • Phagocytic glycoprotein I antibody
    see all

Anti-CD44v6 antibody [VFF-18] images

  • Overlay histogram showing THP1 cells stained with ab78960 (red line). The cells were fixed with 80% methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab78960, 1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.
  • All lanes : Anti-CD44v6 antibody [VFF-18] (ab78960) at 1 µg/ml

    Lane 1 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 2 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 82 kDa
    Observed band size : 91 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 36 kDa,40 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 12 minutes
    CD44v6 contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.
  • ab78960 staining CD44v6 in human breast cancer tissue by Immunohistochemistry (Frozen sections).
    Tissue was fixed in methanol and then blocked using 10% serum for 5 minutes at 25°C. Samples were then incubated with ab78960 at a 1/500 dilution for 1 hour at 25°C. The secondary used was an undiluted HRP conjugated goat polyclonal.

    See Abreview

  • IHC image of ab78960 staining in human breast carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab78960, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

References for Anti-CD44v6 antibody [VFF-18] (ab78960)

This product has been referenced in:
  • Okada T  et al. Coexpression of EpCAM, CD44 Variant Isoforms and Claudin-7 in Anaplastic Thyroid Carcinoma. PLoS One 9:e94487 (2014). IHC-P ; Human . Read more (PubMed: 24727741) »
  • Alamo P  et al. Subcutaneous preconditioning increases invasion and metastatic dissemination in mouse colorectal cancer models. Dis Model Mech 7:387-96 (2014). IHC-P . Read more (PubMed: 24487410) »

See all 3 Publications for this product

Product Wall

The IHC validation of this CD44v6 antibody, ab78960, was carried out on breast carcinoma tissue. We have sections of formalin-fixed, paraffin-embedded ductal breast carcinoma in our catalogue. The catalogue number is ab4697. A literature search suggest...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Sample Human Cell (MKN45)
Specification MKN45
Permeabilization Yes - triton-X 100
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted Aug 02 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (AGS)
Specification AGS
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jul 08 2013

Thank you for your patience while I looked into your question. Ab78960 has not been validated for neutralization studies; however, it has been used in functional assay in Weiss JM et al., The Journal of Cell Biology, Volume 137, 1997. (pubMed ID 916641...

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Thank you for your inquiry. I do not have any information available stating that this antibody can be used for inhibition studies. I will inquire with the antibody originator to see if they have any further information, but it may be that the antibody...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry
Sample Human Cultured Cells (Human breast primary culture)
Specification Human breast primary culture
Fixative Methanol
Blocking step Serum as blocking agent for 5 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Dr. Rene Villadsen

Verified customer

Submitted Jun 23 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Human Tissue sections (Breast cancer)
Specification Breast cancer
Fixative Methanol
Blocking step Serum as blocking agent for 5 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Dr. Rene Villadsen

Verified customer

Submitted Jun 23 2011

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