R-Spondin-1 (RSPO1) is the prototype member of the R-Spondin (RSPO) protein subfamily of a superfamily of thrombospondin type 1 repeat (TSR-1)-containing proteins (Chen et al.; Kamata et al.; Kazanskaya et al.; Kim et al.). Although unable to initialize signaling, RSPO family members are potent enhancers of WNT signaling (Cruciat & Niehrs; de Lau et al.; Kamata et al.; Kazanskaya et al.). They are characterized by a TSR-1 domain, a carboxy-terminal region with positively charged amino acids, and two N-terminal furin-like cysteine-rich repeats (Glinka et al.; Kazanskaya et al.). R-Spondin-1 activates β-catenin signaling via the WNT signaling cascade and by indirectly increasing low-density lipoprotein receptor-related protein 6 (LRP6) on the cell surface. It does this by binding leucine-rich repeat-containing G-protein-coupled receptor 5 (LGR5), and competing with WNT antagonist DKK1 for binding to the WNT coreceptors, Kremen and LRP6, which reduces DKK1-mediated internalization of LRP6 (Binnerts et al.). RSPO1 is involved in a wide range of pleiotropic roles during embryogenesis, it is required for the specification of hematopoietic stem cells, and it has been shown to be important in the growth, survival, and migration of ovarian cancer cells (Cruciat & Niehrs; de Lau et al.; Genthe & Clements; Liu et al.).
(A) The biological activity of Human Recombinant R-Spondin-1 was tested by its ability to induce luciferase activity in a WNT reporter HEK-293 cell line in the presence of mouse Wnt-3a. Luciferase activity was measured using a luminometric assay method. The EC50 is defined as the effective concentration of the growth factor at which luciferase activity is at 50% of maximum. The EC50 in the above example is 36.4 ng/mL.
(B) Human Recombinant R-Spondin-1 was resolved with SDS-PAGE under reducing (+) conditions and non-reducing (-) conditions and visualized by Coomassie Blue staining. Human Recombinant R-Spondin-1 has a predicted molecular mass of 25.6 kDa.
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For differentiation of human ES or iPS cells into hematopoietic progenitor cells
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Human Recombinant R-Spondin-1
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