The Glypicans (glypiated proteoglycans) are a small multigene family of GPI-linked proteoglycans that play a key role in growth factor signaling (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 1 (GPC-1) is synthesized as a 558 amino acid (aa) preproprecursor that contains a 23 aa signal sequence, a 507 aa mature segment, and a 28 aa C-terminal prosegment (6, 7). There are two potential N-linked and four potential O-linked sites for glycosylation or glycanation. There are potentially two heparan sulfate (HS) modifications on GPC-1 that could contribute to a native molecular weight of approximately 200kD (7, 8, 9). Mature human GPC-1 shares 91% aa identity with mature mouse GPC-1. There are two potential splice variants of human GPC-1. Both show an alternate start site at Met73, while one has an additional 65 aa substitution for the C-terminal 264 amino acids (10, 11). Cells known to express GPC-1 include neurons, smooth and skeletal muscle cells, keratinocytes, osteoblasts, Schwann cells, immature dendritic cells, and tumor, plus tumor-associated vascular endothelial cells (8, 9, 12-15). The function of GPC-1 is complex and varied. As a proteoglycan, it appears to make use of its HS adduct to impact select growth factor activity (16). This is accomplished by having juxtramembrane HS attachment sites, and a flexible, GPI-linkage (17). Recent data suggests GPC-1 and sulfation enzymes may collaborate to regulate FGF signaling. HS modules that are rich in 2-O- and 6-O- sulfate upregulate FGF-2 activation of FGFR1c (18). Similarly, FGF-1 requires both 2-O- and 6-O-sulfation to bind to FGFR2c and 3c. By contract, FGF-1 requires no sulfation to bind to FGFR2b, and FGF-8b needs only 6-O-sulfation to activate FGFR3c. Thus, many FGF receptor isoform specific effects may be attributed to an interaction between Glypican family members and the cell sulfation system (19).
Source: A DNA sequence encoding the mature human Glypican 1 (Asp 24-Ser 530; Accession # P35052) was fused at the C-terminus to a six-histidine tag. The protein was expressed in a mouse myeloma cell line, NS0.
Definition: Measured by its ability to bind FGF-basic in a functional ELISA.
Applications:Suitable for use in Western Blot and ELISA. Other applications not tested.
Recommended Dilution:Western Blot: 56.8kD protein migrates as an ~ 60kD protein under reducing conditions
ELISA: 1 at 3ug/ml (100ul/well) will bind rhFGF-basic with an apparent KD ≤1 nM
Optimal dilutions to be determined by the researcher.
Storage and Stability:Lyophilized powder may be stored at 4°C for short-term only. Reconstitute to nominal volume by adding sterile PBS and store at -20°C. Reconstituted product is stable for 12 months at -20°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.
仕様
Size：50ug
Source Antigen：Recombinant Human
Grade：Highly Purified
Purity：≥ 95%, as determined by SDS-PAGE under reducing conditions and visualized by silver stain.Endotoxin ≤ 1EU.
Form：Supplied as a lyophilized powder in PBS. Reconstitute with sterile PBS to ≥ 0.1mg/ml.
