Recombinant human R-spondin 3 protein (Qk032)£155.00 – £1,900.00View product
Recombinant human R-spondin 3 protein (Qk032)
£155.00 – £1,900.00
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.
Qk032 is a 17 kDa 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 origin-free and carrier-protein free.
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£155.00 – £1,900.00
1000µg will be despatched as 2 x 500µg
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Summary:
- High purity bioactive domain of human R-spondin 3 protein (Uniprot: Q9BXY4)
- 17 kDa
>98%, by SDS-PAGE quantitative densitometry
Expressed in E. coli
Animal origin-free (AOF) and carrier protein-free
Manufactured in our Cambridge, UK laboratories
Lyophilized from acetonitrile, TFA
- Resuspend in 10 mM HCl (Reconstitution solution A) at >50 µg/ml, add carrier protein if desired, prepare single-use aliquots and store frozen at -20 °C (short-term) or -80 °C (long-term)
Featured applications:
Chemically defined media for organoid culture
Intestinal organoid culture
Tumor organoid culture
Frequently used together
Alternative proteins
R-spondin 3 activity was determined using the Wnt-responsive firefly luciferase reporter assay. HEK293T cells transfected with reporter TOP-FLASH were treated in triplicate with increasing concentrations of R-spondin 3 in the presence of Wnt-conditioned media (1:8 dilution). RSP1 enhances Wnt-ß catenin signaling in HEK239T cells. Cells were grown overnight, and luciferase activity is measured and normalized. EC50 = 0.3 nM (5 ng/ml). 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.
Further quality assays
Mass spectrometry: single species with expected mass
Recovery from stock vial:Â >95%
Endotoxin: <0.005 EU/μg protein (below level of detection)
We are a company founded and run by scientists to provide a service and support innovation in stem cell biology and regenerative medicine. All our products are exceptionally high purity, with complete characterisation and bioactivity analysis on every lot.
Protein background
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) 4-6 [2]. 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 organoids [2]. 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 labs [3].
Additional resources
Publications using Recombinant human R-spondin 3 protein (Qk032)
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Mass Generation and Long-term Expansion of Hepatobiliary Organoids from Adult Primary Human Hepatocytes
Marsee A, Ritchie A, Myszczyszyn A et al.
DOI: https://doi.org/10.1101/2024.06.10.598262
Our products are for research use only and not for diagnostic or therapeutic use. Products are not for resale.
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