Product Specifications for Recombinant Human Glypican 6 Protein, CF
Mouse myeloma cell line, NS0-derived human Glypican 6 protein Asp24-Arg354 & Ser355-Val527, with a C-terminal 6-His tag
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
<0.10 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis
Asp24 & Ser355
Predicted Molecular Mass
38 kDa and 20 kDa
38 kDa and 27-29 kDa, reducing conditions
Measured by its binding ability in a functional ELISA. When biotinylated recombinant mouse Wnt-3a (25 ng/mL, 100 μL/well) is captured on streptavidin-coated plate, Recombinant Human Glypican 6 binds with an apparent Kd <10 nM.
Formulation, Preparation and Storage
What does CF mean?
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our
Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant
protein to be stored at a more dilute concentration.
The carrier free version does not contain BSA.
What formulation is right for me?
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or
as an ELISA standard.
In contrast, the carrier free protein is recommended for applications, in which the presence of BSA
Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitute at 100 μg/mL in sterile PBS.
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 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 - 70 kDa, 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 110 kDa in size. This translates into approximately 50 kDa 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).
Song, H.H. and J. Filmus (2002) Biochim. Biophys. Acta 1573:241.
Filmus, J. (2001) Glycobiology 11:19R.
De Cat, B. and G. David (2001) Semin. Cell Dev. Biol. 12:117.
Filmus, J. (2003) Glycoconj. J. 19:319.
Veugelers, M. et al. (1999) J. Biol. Chem. 274:26968.
Paine-Saunders, S. et al. (1999) Genomics 57:455.
Fransson, L-A. et al. (2004) Cell Mol. Life Sci. 61:1016.
Fransson, L-A. (2003) Int. J. Biochem. Cell Biol. 35:125.
Bassett, J.H.D. et al. (2006) Endocrinology 147:295.
Which Brands are Currently Available on bio‑techne.com? R&D Systems, Tocris Bioscience and ProteinSimple branded products are available to purchase through bio‑techne.com. ProteinSimple branded instruments are available to quote. ACD branded products will be available on bio‑techne.com in the near future. Novus Biologicals branded products are not currently available on bio‑techne.com and can be found at novusbio.com.
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