Home / Glypican 3 / Human Glypican 3 Antibody

Human Glypican 3 Antibody

Catalog #: AF2119
7 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
AF2119
AF2119-SP
Detection of Human Glypican 3 by Western Blot.
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Product Details
Citations (7)
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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 either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
2 µg/mL
See below
ELISA

This antibody functions as an ELISA detection antibody when paired with Mouse Anti-Human Glypican 3 Monoclonal Antibody (Catalog # MAB21191).

This product is intended for assay development on various assay platforms requiring antibody pairs. We recommend the Human Glypican 3 DuoSet ELISA Kit (Catalog # DY2119) for convenient development of a sandwich ELISA or the Human Glypican 3 Quantikine ELISA Kit (Catalog # DGLY30) for  a complete optimized ELISA.

 
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.

Scientific Data

Western Blot Detection of Human Glypican 3 antibody 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 antibody in HepG2 Human Cell Line by Immunocytochemistry (ICC). 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 antibody in Human Breast by Immunohistochemistry (IHC-P). 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 antibody in Human Liver Cancer Tissue by Immunohistochemistry (IHC-P). 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.

ELISA View Larger

Human Glypican 3 ELISA Standard Curve. Recombinant Human Glypican 3 protein was serially diluted 2-fold and captured by Mouse Anti-Human Glypican 3 Monoclonal Antibody (MAB21191) coated on a Clear Polystyrene Microplate (DY990). Sheep Anti-Human Glypican 3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2119) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (DY998) followed by Substrate Solution (DY999) and stopping the enzymatic reaction with Stop Solution (DY994).

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.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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); 102137748 (Cynomolgus Monkey)
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

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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.

7 Citations: Showing 1 - 7
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  1. Purification of HCC-specific extracellular vesicles on nanosubstrates for early HCC detection by digital scoring
    Authors: N Sun, YT Lee, RY Zhang, R Kao, PC Teng, Y Yang, P Yang, JJ Wang, M Smalley, PJ Chen, M Kim, SJ Chou, L Bao, J Wang, X Zhang, D Qi, J Palomique, N Nissen, SB Han, S Sadeghi, RS Finn, S Saab, RW Busuttil, D Markovic, D Elashoff, HH Yu, H Li, AP Heaney, E Posadas, S You, JD Yang, R Pei, VG Agopian, HR Tseng, Y Zhu
    Nat Commun, 2020-09-07;11(1):4489.
    Species: Human
    Sample Types: Plasma
    Applications: ELISA Capture
  2. Bone Marrow-Derived Mesenchymal Stem Cell Potential Regression of Dysplasia Associating Experimental Liver Fibrosis in Albino Rats
    Authors: YH Khalifa, GM Mourad, WM Stephanos, SA Omar, RA Mehanna
    Biomed Res Int, 2019-11-06;2019(0):5376165.
    Species: Rat
    Sample Types: Whole Tissue
    Applications: IHC-P
  3. Expression of Glypican 3 is an Independent Prognostic Biomarker in Primary Gastro-Esophageal Adenocarcinoma and Corresponding Serum Exosomes
    Authors: M Rahbari, M Pecqueux, D Aust, H Stephan, O Tiebel, A Chatzigeor, T Tonn, F Baenke, V Rao, N Ziegler, H Greif, K Lin, J Weitz, NN Rahbari, C Kahlert
    J Clin Med, 2019-05-16;8(5):.
    Species: Human
    Sample Types: Exosome Lysates, Exosomes
    Applications: Flow Cytometry, Western Blot
  4. Regulation of bone morphogenetic protein signalling and cranial osteogenesis by Gpc1 and Gpc3
    Authors: Prem P. Dwivedi, Randall H. Grose, Jorge Filmus, Charles S.T. Hii, Cory J. Xian, Peter J. Anderson et al.
    Bone
  5. 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., 2012-05-10;18(13):3686-96.
    Species: Human
    Sample Types: Serum
    Applications: ELISA Development
  6. The oncofetal protein glypican-3 is a novel marker of hepatic progenitor/oval cells.
    Authors: Grozdanov PN, Yovchev MI, Dabeva MD
    Lab. Invest., 2006-10-02;86(12):1272-84.
    Species: Rat
    Sample Types: Whole Tissue
    Applications: IHC-Fr
  7. 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., 2006-10-01;209(1):219-29.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot

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