Recombinant human R-spondin 3 protein (Qk032)
Human R-spondin 3 protein potentiates Wnt signalling in and has been shown to function in crypt regeneration in the intestine and control stem cell and progenitor cell behaviour during kidney development. R-spondin 3 is used alongside R-spondin 1 in intestinal organoid culture systems.
17kDa highly pure, bioactive domain of human R-spondin 3 comprising the two cysteine-rich furin-like domains, which are necessary and sufficient for Wnt signalling potentiation and are the essential domains for activity in stem cell and organoid culture. Animal-free and carrier-protein free.
Bioactivity using a Wnt reporter assay in HEK293T cells. HEK293T cells transfected with Wnt-responsive firefly luciferase reporter TOPflash are treated with increasing concentrations of R-spondin 3 in the presence of Wnt-conditioned media (1:8 dilution), in triplicate. The following day, luciferase activity is measured by luminescence. EC50 = 4.02 ng/ml (0.3 nM). Data from Qk032 #010
R-spondin 3 bioactive domain migrates at 17 kDa in non-reducing (-βME) conditions and upon reduction (+βME).
Purified recombinant protein (7 µg) was resolved using 15% w/v SDS-PAGE in reduced (+β-mercaptothanol, R) and non-reduced conditions (NR) and stained with Coomassie Brilliant Blue R250.
Data from Qk032 lot #010
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R-Spondin 3 (RSPO3) protein belongs to the R-spondin family comprizing four structurally-related, secreted ligands (RSPO 1-4)1. All contain furin-like and thrombospondin structural domains. R-spondins are activators of the canonical Wnt signaling pathway. Receptors include the leucine-rich repeat-containing G-protein coupled receptors (LGR) 4-62. In the colon, R-spondin 3 promotes stem cell recovery and epithelial repair via induction of Wnt.
An R-spondin, such as R-spondin 3, is an essential component of Wnt 3a, R-spondin-3 (or 1,2), Noggin (WRN) media used widely in propagation of intestinal organoids2. There is a drive towards improved reproducibility in stem cell and organoid culture. Conditioned media (spent media from cells that secrete growth factors such as Wnt and R-spondin, along with several other factors and waste products) is both challenging to manufacture consistently and conveniently, and a source of variability between labs3.
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