Human Glypican 3 Antibody

Catalog # Availability Size / Price Qty
AF2119
AF2119-SP
Human Glypican 3 Antibody in Western Blot
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Product Details
Citations (3)
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Human Glypican 3 Antibody Summary

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

Western Blot Detection of Human Glypican 3 by Western Blot. View Larger

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. View Larger

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. View Larger

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. View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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

Product Datasheets

Citations for Human Glypican 3 Antibody

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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  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 Types: Serum
    Applications: ELISA Development
  2. 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 Types: Cell Lysates
    Applications: Western Blot
  3. 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 Types: Whole Tissue
    Applications: IHC-Fr

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