Recombinant human R-spondin 3 protein (Qk032) Price range: £175.00 through £1,900.00View product
Recombinant human R-spondin 3 protein (Qk032)
Price range: £175.00 through £1,900.00
Human R-spondin 3 protein potentiates Wnt signaling in and has been shown to function in crypt regeneration in the intestine and control stem cell and progenitor cell behavior 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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Price range: £175.00 through £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.
For use in manufacturing of cellular or gene therapy products. Not intended for in vivo applications.
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