Human/Mouse/Rat Glypican 3 Antibody

(1 citations)   
  • Species Reactivity
    Human, Mouse, Rat
  • Specificity
    Detects human Glypican 3 in ELISAs. Deteacts human, mouse, and rat Glypican 3 in Western blots. Does not cross-react with recombinant human (rh) Glypican-2, rhGlypican-5, or rhGlypican-6.
  • Source
    Monoclonal Mouse IgG2A Clone # 307801
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human Glypican 3
    Gln25-Val558
    Accession # P51654.1
  • 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
    1-2 µg/mL
    See below
  • Flow Cytometry
    2.5 µg/106 cells
    HepG2 human hepatocellular carcinoma cell line
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    3-25 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 Mouse Anti-Human Glypican 3 Monoclonal Antibody (Catalog # MAB2119) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). 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.
Detection of Mouse and Rat Glypican 3 by Western Blot. Western blot shows lysates of mouse placenta tissue, mouse lung tissue, mouse adrenal gland tissue, rat placenta tissue, and rat lung tissue. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Glypican 3 Monoclonal Antibody (Catalog # MAB2119) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Glypican 3 at approximately 65-70 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 Mouse Anti-Human Glypican 3 Monoclonal Antibody (Catalog # MAB2119) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) 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.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 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.

1 Citations: Showing 1 - 1

  1. The heparan sulfate proteoglycan (HSPG) glypican-3 mediates commitment of MC3T3-E1 cells toward osteogenesis.
    Authors: Haupt LM, Murali S, Mun FK, Teplyuk N, Mei LF, Stein GS, van Wijnen AJ, Nurcombe V, Cool SM
    J. Cell. Physiol., 2009;220(3):780-91.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
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Mouse IgG2A Isotype Control

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Secondary Antibodies
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Mouse IgG HRP-conjugated Antibody

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Mouse IgG HRP-conjugated Antibody

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Mouse F(ab)2 IgG (H+L) APC-conjugated Antibody

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Donkey Anti-Mouse IgG NorthernLights™ NL637-conjugated Antibody

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Donkey Anti-Mouse IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL009 5
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Mouse F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody

Flow F0103B 5  
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Mouse IgG Antibody

WB AF007 4
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Goat Anti-Mouse IgG Biotinylated Antibody

WB BAF007 4
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Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC002  
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Mouse IgG VisUCyte HRP Polymer Antibody

IHC VC001  
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Rat Anti-Mouse IgG2A PE-conjugated Antibody

Flow F0129 1  
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Goat Anti-Mouse IgG Fc Affinity Purified Polyclonal Ab

G-202-C 2
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Mouse F(ab)2 IgG (H+L) PerCP-conjugated Antibody

Flow F0114  
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Rat Anti-Mouse IgG2A APC-conjugated Antibody

Flow F0130  
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Mouse IgG2A Antibody

Flow MAB0032  
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Rat Anti-Mouse IgG2A PerCP-conjugated Antibody

Flow F0131  
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