Glypican 6, Recombinant, Human (Glypiated Proteoglycan 6)

The Glypicans (glypiated proteoglycans) are a small multigene family of GPI-linked heparan sulfate (HS) proteoglycans that likely play a key role in embryonic morphogenesis (1, 2, 3, 4). There are currently six known mammalian Glypicans. They all share a common-sized protein core of 60-70kD, an N-terminus which likely forms a compact globular domain, 14 conserved cysteines that form multiple intrachain disulfide bonds, and a number of C-terminal N- and O-linked carbohydrate attachment sites. Based on exon organization and the location of O-linked glycosylation sites, at least two subfamilies of Glypicans are known, with one subfamily containing Glypicans 1, 2, 4 and 6, and another subfamily containing Glypicans 3 and 5 (3, 5). Human Glypican 6 (GPC-6) is synthesized as a 554 amino acid (aa) preproprecursor that contains a 23 aa signal sequence, a 505 aa mature region and a 26 aa C-terminal prosegment (5, 6). There are four consecutive Ser-Gly repeats that serve as a heparin sulfate attachment site. GPC-6 is reported to be as large as 110kD in size. This translates into approximately 50kD of proteoglycan (5). Human to mouse, there is 97% aa identity over the entire GPC-6 molecule. Cells known to express GPC-6 are adult ovary and embryonic vascular and visceral smooth muscle, plus mesenchyme (embryonic connective tissue) in multiple organs (1, 5, 6). The function of GPC-6 is essentially unknown. As a Glypican family member, it may facilitate heparin-binding growth factor signaling and polyamine uptake into expressing cells (7, 8). In this regard, it would appear that GPC-6 with its attendant HS is downregulated by triiodothyronine during cartilage maturation, thus limiting the availability of sites for FGF sequestration and activity (9).\n\nSource: DNA sequence encoding the mature human Glypican 6 (Asp 24-Val 527; Accession # Q9Y625) was fused with six Histidines at the C-terminus and the protein was expressed in a mouse myeloma cell line, NS0.\n\nDefinition: Measured by its ability to bind biotinylated rmWnt-3A captured on streptavidin-coated plate.\n\nApplications: \nSuitable for use in Western Blot. Other applications not tested.\n\nRecommended Dilution:\nWestern Blot: 38kD and 20kD, respectively migrates at ~ 38 and 27-29kD under reducing conditions. \nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nLyophilized powder may be stored at 4 degrees C for short-term only. Reconstitute to nominal volume by adding sterile PBS and store at -20 degrees C. Reconstituted product is stable for 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

G8235-58H

Size

50ug

Applications

WB

Reactivities

Hum

Form

Supplied as a lyophilized powder in PBS. Reconstitute with sterile PBS to (same/more than) 0.1mg/ml.

Purity

(same/more than) 95%, as determined by SDS-PAGE under reducing conditions and visualized by silver stain. Endotoxin (same/less than) 1EU.

References

1. Song, H.H. & J. Filmus (2002) Biochim. Biophys. Acta 1573:241. 6. Paine-Saunders, S. et al. (1999) Genomics 57:455.\n2. Filmus, J. (2001) Glycobiology 11:19R. 7. Fransson, L-A. et al. (2004) Cell Mol. Life Sci. 61:1016.\n3. De Cat, B. & G. David (2001) Semin. Cell Dev. Biol. 12:117. 8. Fransson, L-A. (2003) Int. J. Biochem. Cell Biol. 35:125.\n4. Filmus, J. (2003) Glycoconj. J. 19:319. 9. Bassett, J.H.D. et al. (2006) Endocrinology 147:295.\n5. Veugelers, M. et al. (1999) J. Biol. Chem. 274:26968.