Recombinant Human Glypican 3 Protein, CF

Recombinant human Glypican 3 (2119-GP/CF) binds recombinant human FGF basic 146 in a functional ELISA. The concentration of recombinant human FGF basic 146 that produces 50% of the optimal binding response is 0.6-3 ng/mL.

• Reactivity: Hu
• Applications: Binding Activity
• Format: Carrier-Free

Recombinant Human Glypican 3 Protein, CF Summary

• Details of Functionality: Measured by its binding ability in a functional ELISA. When Recombinant Human Glypican 3 is immobilized at 0.5 μg/mL (100 µL/well), the concentration of Recombinant Human FGF basic 146 aa (Catalog # 233-FB) that produces 50% of the optimal binding response is approximately 0.6-3 ng/mL.
• Source: Mouse myeloma cell line, NS0-derived human Glypican 3 protein
  Met1-His559 with a C-terminal 6-His tag
• Accession #: P51654.1
• N-terminal Sequence: No results obtained: Gln25 predicted, Ser359 & Val483
• Structure / Form: Glypican 3 is subject to endoproteolytic processing by proprotein convertases (PC). By amino acid sequencing, three peptides (the first with a blocked N-terminus most likely starts with Gln25, the second peptide starts with Ser359 after a furin cleavage site, and the third peptide starts with Val483) are present in the recombinant GPC3 preparation. Peptides 2 and 3 are detected at a 1:1 ratio. All three peptides remained associated via disulfide bonds.
• Protein/Peptide Type: Recombinant Proteins
• Gene: GPC3
• Purity: >97%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Binding Activity
• Theoretical MW: 61.6 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 60-100 kDa, non-reducing conditions

Packaging, Storage & Formulations

• 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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS.
• Purity: >97%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Reconstitution Instructions: Reconstitute at 100 μg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human Glypican 3 Protein, CF

• 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

Background

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).

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.

Publications for Glypican 3 (2119-GP/CF)(7)

Publications using 2119-GP/CF

• TL Hickman, E Choi, KR Whiteman, S Muralidhar, T Pai, T Johnson, A Parikh, T Friedman, M Gilbert, B Shen, L Barron, KE McGinness, SA Ettenberg, GT Motz, GJ Weiss, A Jensen-Smi BOXR1030, an anti-GPC3 CAR with exogenous GOT2 expression, shows enhanced T cell metabolism and improved anti-cell line derived tumor xenograft activity PLoS ONE, 2022-05-04;17(5):e0266980. 2022-05-04 [PMID: 35507536] (Bioassay, Human)
• M Miura, N Fujinami, Y Shimizu, S Mizuno, K Saito, T Suzuki, M Konishi, S Takahashi, N Gotohda, K Suto, T Yoshida, T Nakatsura Usefulness of plasma full-length glypican-3 as a predictive marker of hepatocellular carcinoma recurrence after radial surgery Oncol Lett, 2020-02-05;19(4):2657-2666. 2020-02-05 [PMID: 32218816] (ELISA Development, Human)
• T Ueda, A Kumagai, S Iriguchi, Y Yasui, T Miyasaka, K Nakagoshi, K Nakane, K Saito, M Takahashi, A Sasaki, S Yoshida, N Takasu, H Seno, Y Uemura, K Tamada, T Nakatsura, S Kaneko Non-clinical efficacy, safety and stable clinical cell processing of induced pluripotent stem cell-derived anti-glypican-3 chimeric antigen receptor-expressing natural killer/innate lymphoid cells Cancer Sci., 2020-03-31;111(5):1478-1490. 2020-03-31 [PMID: 32133731] (In Vivo, Mouse)
• Epitope mapping by a Wnt-blocking antibody: evidence of the Wnt binding domain in heparan sulfate Sci Rep, 2016-05-17;6(0):26245. 2016-05-17 [PMID: 27185050] (ELISA (Standard), Human)
• 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. 2012-05-10 [PMID: 22577059] (ELISA (Standard))
• Komori H, Nakatsura T, Senju S, Yoshitake Y, Motomura Y, Ikuta Y, Fukuma D, Yokomine K, Harao M, Beppu T, Matsui M, Torigoe T, Sato N, Baba H, Nishimura Y Identification of HLA-A2- or HLA-A24-restricted CTL epitopes possibly useful for glypican-3-specific immunotherapy of hepatocellular carcinoma. Clin. Cancer Res., 2006-05-01;12(9):2689-97. 2006-05-01 [PMID: 16675560] (ELISA (Standard))
• Motomura Y, Senju S, Nakatsura T, Matsuyoshi H, Hirata S, Monji M, Komori H, Fukuma D, Baba H, Nishimura Y Embryonic stem cell-derived dendritic cells expressing glypican-3, a recently identified oncofetal antigen, induce protective immunity against highly metastatic mouse melanoma, B16-F10. Cancer Res., 2006-02-15;66(4):2414-22. 2006-02-15 [PMID: 16489048] (Bioassay, Mouse)

Bioinformatics

• Gene Symbol: GPC3
• Uniprot:
  • P51654.1()