Recombinant Mouse Glypican 3 Protein, CF

R&D Systems, part of Bio-Techne | Catalog # 6938-GP

R&D Systems, part of Bio-Techne
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6938-GP-050
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Key Product Details

Source

NS0

Accession #

AAH36126

Conjugate

Unconjugated

Applications

Bioactivity

View all Glypican 3 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse Glypican 3 protein
Gln25-Met557 & Ser358-Met557, both with a C-terminal 6-His tag

Purity

>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

 Gln25 predicted (No result obtained, sequencing might be blocked) & Ser358

Predicted Molecular Mass

61.3 kDa

SDS-PAGE

66-98 kDa & 30-40 kDa, reducing conditions

Activity

Measured by its binding ability in a functional ELISA.
When Recombinant Mouse Glypican 3 is coated at 5 μg/mL (100 μL/well), the concentration of recombinant human FGF basic that produces 50% of the optimal binding response is found to be approximately 0.75-3.75 ng/mL.

Formulation, Preparation and Storage

6938-GP
Formulation Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution
Reconstitute at 100 μg/mL in PBS.

Reconstitution Buffer Available:
Size / Price
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Shipping The product is shipped at ambient temperature. Upon receipt, store it 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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Glypican 3

The Glypicans (GPCs) are a small multigene family of GPI-linked proteoglycans that play a key role in growth factor signaling (1 ‑ 4). Glypicans exhibit a 60 ‑ 70 kDa protein core that consists of a globular N‑terminus, 14 conserved cysteines that form multiple intrachain disulfide bonds, and a number of C‑terminal N‑ and O‑linked carbohydrate attachment sites. At least two subfamilies of Glypicans are known (GPC1, 2, 4, and 6 and GPC3 and 5) (1, 5). Mouse Glypican 3 is synthesized as a 579 amino acid (aa) preproprecursor that contains a 24 aa signal sequence, a 535 aa mature segment, and a 20 aa C-terminal prosegment (6 ‑ 8). There are three potential N-linked, and two potential O-linked sites for glycosylation or glycanation. O-linked glycanation utilizes heparan sulfate (HS) and this contributes 60 ‑ 120 kDa to an overall molecular weight of approximately 200 kDa for Glypican 3 (9, 10). Glypican 3 undergoes convertase‑mediated proteolytic processing between Arg357 ‑ Ser358 to generate a disulfide-linked cell-surface heterodimer (9). It may also undergo Notum cleavage of the GPI-anchor to create a soluble circulating form (11, 12). Mature mouse Glypican 3 shares 95% and 99% aa identity with mature human and rat Glypican 3, respectively. Sources for Glypican 3 include liver carcinoma cells plus mesoderm and mesodermal derivatives. Cell types in mice include mesenchyme, prechondrocytes, primitive hematopoietic progenotors, fetal hepatocytes, and pericardium (6, 13). Glypican 3 is best known in human for its association with SGB syndrome, a condition characterized by tissue overgrowth (dysmorphogenesis), potentially through its binding to CD26. Normally, Glypican 3 binds to and down-regulates cell surface CD26.  In the absence of Glypican 3, CD26 promotes cell proliferation, possibly resulting in tissue malformation(s) (14 ‑ 16). Glypican 3 makes use of its both its core protein and HS adduct to impact select growth factor activity. The core protein is known to bind to Wnt and stabilize the Wnt:Frizzled interactions (1, 17, 18). It also binds to hedgehog (HH), serving as a competitive inhibitor for Patched. This results in the internalization and degradation of circulating HH, and it also removes a soluble promoter of Smoothened signaling (1, 19). The HS component of Glypican 3, by contrast, is reported to interact with members of the FGF system, promoting FGF receptor dimerization and signaling (1, 2). Finally, Glypican 3 binds to surface membrane Glut4, facilitating its stabilization and promoting the transport of glucose into the cell (20).

References

  1. Filmis, J. et al. (2008) Genome Biol. 9:224.
  2. Fransson, L-A. et al. (2004) Cell. Mol. Life Sci. 61:1016.
  3. Fransson, L-A. (2003) Int. J. Biochem. Cell Biol. 35:125.
  4. Lamoureux, F. et al. (2007) BioEssays 29:758.
  5. Veugelers, M. et al. (1999) J. Biol. Chem. 274:26968.
  6. Pellegrini, M. et al. (1998) Dev. Dyn. 213:431.
  7. Watanabe, K. et al. (1995) J. Cell Biol. 130:1207.
  8. Ho, M. and H. Kim (2011) Eur. J. Cancer 47:333.
  9. Capurro, M.I. et al. (2005) J. Biol. Chem. 280:41201.
  10. Filmus, J. et al. (1995) Biochem. J. 311:561.
  11. Traister, A. et al. (2008) Biochem. J. 410:503.
  12. Liu, H. et al. (2010) World J. Gastroenterol. 16:4410.
  13. Viviano, B.L. et al. (2005) Dev. Biol. 282:152.
  14. Gonzales, A.D. et al. (1998) J. Cell Biol. 141:1407.
  15. Cano-Gauci, D.F. et al. (1999) J. Cell Biol. 146:255.
  16. Cavoodi, J. et al. (2007) Proteomics 7:2300.
  17. Song, H.H. et al. (2005) J. Biol. Chem. 280:2116.
  18. Capurro, M.I. et al. (2005) Cancer Res. 65:6245.
  19. Capurro, M.I. et al. (2008) Dev. Cell 14:700.
  20. Taguchi, A. et al. (2008) Biochem. Biophys. Res. Commun. 369:1204.

Alternate Names

GPC3

Entrez Gene IDs

2719 (Human); 14734 (Mouse); 25236 (Rat); 102137748 (Cynomolgus Monkey)

Gene Symbol

GPC3

UniProt

AAH36126

Additional Glypican 3 Products

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