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Active Glypican 1 (GPC1)

Glypican Proteoglycan 1; Secreted glypican-1

ACTIVITY TEST

Glypican 1 (GPC1) is one of the six glycosylphosphatidylinositol-anchored, cell surface heparan sulfate proteoglycans that acts as a growth factor signaling. 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. The function of GPC-1 is complex and varied, it can modulate various signaling pathways including Heparin Binding Epidermal Growth Factor Like Growth Factor (HBEGF), fibroblast growth factors (FGF), vascular endothelial growth factor-A (VEGF-A), transforming growth factor-β (TGF-β), Wnt, Hedgehog (Hh), and bone morphogenic protein (BMP) through specific interactions with pathway ligands and receptors. Thus a functional binding ELISA assay was conducted to detect the interaction of recombinant human GPC1 and recombinant human HBEGF. Briefly, GPC1 was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to HBEGF-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-GPC1 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody for 1h at 37℃, wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50 µL stop solution to the wells and read at 450/630 nm immediately. The binding activity of recombinant human GPC1 and recombinant human HBEGF was shown in Figure 1, the EC50 for this effect is 0.17 ug/mL.

USAGE

Reconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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Magazine Citations
Intensive Care Medicine Experimental Elevated plasma glypicans are associated with organ failure in patients with infection Pubmed: 30618011
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