Human Glypican 3 Antibody

(3 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human Glypican 3 in direct ELISAs and Western blots. In direct ELISAs, less than 10% cross-reactivity with recombinant mouse Glypican 3 is observed and less than 5% cross-reactivity with recombinant human (rh) Glypican 2, rhGlypican 5, and rhGlypican 6 is observed.
  • Source
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human Glypican 3
    Gln25-Val558
    Accession # P51654
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    2 µg/mL
    See below
  • Flow Cytometry
    2.5 µg/106 cells
    HepG2 human hepatocellular carcinoma cell line
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    1-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human Glypican 3 by Western Blot. Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line. PVDF membrane was probed with 2 µg/mL of Sheep Anti-Human Glypican 3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2119) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Glypican 3 at approximately 75 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
Glypican 3 in HepG2 Human Cell Line. Glypican 3 was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line using Sheep Anti-Human Glypican 3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2119) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and cell membranes. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Immunohistochemistry
Glypican 3 in Human Breast. Glypican 3 was detected in immersion fixed paraffin-embedded sections of human breast using 5 µg/mL Sheep Anti-Human Glypican 3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2119) overnight at 4 °C. Tissue was stained with the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Immunohistochemistry
Glypican 3 in Human Liver Cancer Tissue. Glypican 3 was detected in immersion fixed paraffin-embedded sections of human liver cancer tissue using Sheep Anti-Human Glypican 3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2119) at 10 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Glypican 3

Glypicans (GPC) are a family of heparan sulfate proteoglycans that are attached to the cell surface by a glycosylphosphatidylinositol (GPI) anchor. Six members of this family have been identified in mammals (GPC1-GPC6). All glypican core proteins contain an N-terminal signal peptide, a large globular cysteine-rich domain (CRD) with 14 invariant cysteine residues, a stalk-like region containing the heparan sulfate attachment sites, and a C-terminal GPI attachment site. While glypican proteins do not share strong amino acid sequence identity (they range from 17-63%), the conserved cysteine residues in their CRDs suggests similarity in their three‑dimensional structure (1, 2).

Mutations in GPC3 cause a rare disorder in humans, Simpson-Golabi-Behmel Syndrome, which is characterized by pre and postnatal overgrowth of multiple tissues and organs and an increased risk for developing embryonic tumors (3). These features are also present in the mouse knock-out of GPC3 indicating that GPC3 regulates cell survival and inhibits cell proliferation during development (4). Glypican 3 has been implicated in regulating many different signaling pathways including: IGF, FGF, BMP, and Wnt. An endoproteolytic processing of GPC3 by proprotein convertases is required for the modulation of Wnt signaling (5). Direct interaction with FGF-basic has been observed and is mediated by the heparan sulfate chains (6).

  • References:
    1. Filmus, J. and S.B. Selleck (2001) J. Clinical Invest. 108:497. 
    2. De Cat, B and G. David (2001) Seminars in Cell & Dev. Biol. 12:117. 
    3. Pilia, G. et al. (1996) Nat. Genet. 12: 241. 
    4. Cano-Gauci, D.F. et al. (1999) J. Cell Biol. 146: 255.
    5. De Cat, B. et al. (2003) J. Cell Biol. 163:625.
    6. Song, H.H. et al. (1997) J. Biol. Chem. 272:7574.
  • Entrez Gene IDs:
    2719 (Human); 14734 (Mouse); 25236 (Rat)
  • Alternate Names:
    DGSX; Glypican 3; glypican proteoglycan 3; glypican-3; GPC3; GTR2-2; heparan sulphate proteoglycan; Intestinal protein OCI-5; MXR7; OCI5; OCI-5; secreted glypican-3; SGB; SGBS; SGBS1SDYS
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

3 Citations: Showing 1 - 3
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Species
Applications
Sample Type
  1. Phase I trial of a glypican-3-derived peptide vaccine for advanced hepatocellular carcinoma: immunologic evidence and potential for improving overall survival.
    Clin. Cancer Res., 2012;18(13):3686-96.
    Species: Human
    Sample Type: Serum
    Application: ELISA Development
  2. The oncofetal protein glypican-3 is a novel marker of hepatic progenitor/oval cells.
    Authors: Grozdanov PN, Yovchev MI, Dabeva MD
    Lab. Invest., 2006;86(12):1272-84.
    Species: Rat
    Sample Type: Whole Tissue
    Application: IHC Frozen
  3. Bone-specific heparan sulfates induce osteoblast growth arrest and downregulation of retinoblastoma protein.
    Authors: Manton KJ, Sadasivam M, Cool SM, Nurcombe V
    J. Cell. Physiol., 2006;209(1):219-29.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
Cell and Tissue Staining Kits
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Anti-Sheep HRP-DAB Cell & Tissue Staining Kit

CTS019 1
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Immunohistochemistry Reagents
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Antigen Retrieval Reagent-Universal

CTS015 10
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Antigen Retrieval Reagent-Basic

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Mounting Medium

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Antigen Retrieval Reagent Sampler (50 mL each)

CTS016
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Antigen Retrieval Reagent-Acidic

CTS014
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NorthernLights Guard Mounting Media

NL996
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VisUCyte Antigen Retrieval Reagent-Acidic

VCTS022
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VisUCyte Antigen Retrieval Reagent-Basic

VCTS021
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VisUCyte Antigen Retrieval Reagent-Universal

VCTS023
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Isotype Controls
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Normal Sheep IgG Control

Ctrl 5-001-A 9
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Secondary Antibodies
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Sheep IgG HRP-conjugated Antibody

WB, Simple Western HAF016 14  
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Donkey Anti-Sheep IgG NorthernLights™ NL557-conjugated Antibody

Flow, IHC, ICC NL010 3
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Donkey Anti-Sheep IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL011 5
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Donkey Anti-Sheep IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL012 2
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Donkey Anti-Sheep IgG Biotinylated Antibody

WB BAF016 2
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Sheep IgG (H+L) PE-conjugated Antibody

Flow F0126  
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Sheep IgG (H+L) APC-conjugated Antibody

Flow F0127  
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Sheep IgG VisUCyte HRP Polymer Antibody

IHC VC006  
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Donkey Anti-Sheep IgG (H+L) Fluorescein-conjugated Antibody

Flow F0125 1
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Donkey Anti-Sheep IgG (H+L) PerCP-conjugated Antibody

Flow F0128
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