Recombinant Mouse R-Spondin 3 Protein

• Reactivity: Mu
• Applications: Bioactivity

Recombinant Mouse R-Spondin 3 Protein Summary

• Details of Functionality: Measured by its ability to induce Topflash reporter activity in HEK293T human embryonic kidney cells. The ED₅₀ for this effect is 0.25-1 ng/mL in the presence of 5 ng/mL Recombinant Mouse Wnt‑3a (Catalog # 1324-WN).
• Source: Chinese Hamster Ovary cell line, CHO-derived mouse R-Spondin 3 protein
  Met1-His277
• Accession #: Q2TJ95
• N-terminal Sequence: Met33
• Protein/Peptide Type: Recombinant Proteins
• Gene: Rspo3
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 28.9 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 37-43 kDa, reducing conditions

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Reconstitution Instructions: Reconstitute at 200 μg/mL in PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Mouse R-Spondin 3 Protein

• Cristin 1
• CRISTIN1
• FLJ14440
• hPWTSR
• hRspo3
• Protein with TSP type-1 repeat
• PWTSR
• Roof plate-specific spondin-3
• RSPO3
• R-spondin 3 homolog (Xenopus laevis)
• RSpondin 3
• R-Spondin 3
• R-spondin-3
• Thrombospondin type-1 domain-containing protein 2
• thrombospondin, type I, domain containing 2
• THSD2

Background

R-Spondin 3 (RSPO3, roof plate-specific spondin 3), also called cysteine-rich and single thrombospondin domain containing-1 (Cristin 1), is an ~31 kDa secreted protein that shares ~40% aa identity with the other three R‑Spondin family members (1, 2). All are positive modulators of Wnt/ beta -catenin signaling, but each has a distinct expression pattern (1-4). Like other R‑spondins, R‑Spondin 3 contains two adjacent cysteine-rich furin‑like domains (amino acids (aa) 35-135) with one potential N‑glycosylation site (aa 36), followed by a thrombospondin (TSP-1) motif (aa 147-207) and a region rich in basic residues (aa 211-269). Only the furin-like domains are needed for beta -catenin stabilization (2). Within aa 21-209, mouse R‑Spondin 3 shares 93%, 97%, 96%, 95% and 91% aa identity with human, rat, equine, bovine and canine R‑Spondin 3, respectively. Potential isoforms of 217, 224 and 252 aa are divergent or truncated at the C terminus; the 252 aa form also lacks aa 4‑33 at the N‑terminus (5). Mouse R‑Spondin 3 is critical for development of the placental labyrinthine layer, probably by promoting VEGF expression and thus vascular development (6, 7). It is also essential for expression of the placenta-specific transcription factor, Gcm1. In the mouse embryo, R‑Spondin 3 is often expressed by or located near endothelial cells (6). It is found in the roof plate, tail, somites, otic vesicles, cephalic mesoderm, truncus arteriosus, atrioventricular canal of the developing heart, and strongly but transiently in developing limbs (4, 7). R‑Spondins regulate Wnt/ beta -catenin by competing with the Wnt antagonist DKK-1 for binding to the Wnt co-receptors LRP-6 and Kremen, reducing their DKK‑1‑mediated internalization (8, 9). Reports differ on whether R‑Spondins bind LRP-6 directly (8-10). R‑Spondin 3 has also been identified as an oncogene (11).

1. Chen, J-Z. et al. (2002) Mol. Biol. Rep. 29:287.
2. Kim, K.-A. et al. (2008) Mol. Biol. Cell 19:2588.
3. Hendrickx, M. and L. Leyns (2008) Develop. Growth Differ. 50:229.
4. Nam, J.-S. et al. (2007) Gene Expr. Patterns 7:306.
5. Entrez Accession # BAB28811, BAC36296 and EDL04842.
6. Kazanskaya, O. et al. (2008) Development 135:3655.
7. Aoki, M. et al. (2007) Dev. Biol. 301:218.
8. Binnerts, M.E. et al. (2007) Proc. Natl. Acad. Sci. USA 104:14700.
9. Nam, J.-S. et al. (2006) J. Biol. Chem. 281:13247.
10. Wei, Q. et al. (2007) J. Biol. Chem. 282:15903.
11. Theodorou, V. et al. (2007) Nat. Genet. 6:759.

Publications for R-Spondin 3 (4120-RS)(13)

Publications using 4120-RS

• K Nagano, K Yamana, H Saito, R Kiviranta, AC Pedroni, D Raval, C Niehrs, F Gori, R Baron R-spondin 3 deletion induces Erk phosphorylation to enhance Wnt signaling and promote bone formation in the appendicular skeleton Elife, 2022-11-02;11(0):. 2022-11-02 [PMID: 36321691] (Bioassay, Mouse)
• Y Mita, H Zhu, Y Furuichi, H Hamaguchi, Y Manabe, NL Fujii R-spondin3 is a myokine that differentiates myoblasts to type I fibres Scientific Reports, 2022-07-29;12(1):13020. 2022-07-29 [PMID: 35906363] (Bioassay, Mouse)
• KH Nilsson, P Henning, ME Shahawy, M Nethander, TL Andersen, C Ejersted, J Wu, KL Gustafsson, A Koskela, J Tuukkanen, PPC Souza, J Tuckermann, M Lorentzon, LE Ruud, T Lehtimäki, JH Tobias, S Zhou, UH Lerner, JB Richards, S Movérare-S, C Ohlsson RSPO3 is important for trabecular bone and fracture risk in mice and humans Nature Communications, 2021-08-13;12(1):4923. 2021-08-13 [PMID: 34389713] (Bioassay, Mouse)
• M Zhang, M Haughey, NY Wang, K Blease, AM Kapoun, S Couto, I Belka, T Hoey, M Groza, J Hartke, B Bennett, J Cain, A Gurney, B Benish, P Castiglion, C Drew, J Lachowicz, L Carayannop, SD Nathan, J Distler, DA Brenner, K Hariharan, H Cho, W Xie Targeting the Wnt signaling pathway through R-spondin 3 identifies an anti-fibrosis treatment strategy for multiple organs PLoS ONE, 2020-03-11;15(3):e0229445. 2020-03-11 [PMID: 32160239] (Cell Culture, IHC Control, Mouse)
• KK Dickinson, LC Hammond, CM Karner, ND Hastie, TJ Carroll, P Goodyer Molecular determinants of WNT9b responsiveness in nephron progenitor cells PLoS ONE, 2019-04-12;14(4):e0215139. 2019-04-12 [PMID: 30978219] (Bioassay, Mouse)
• R Ogasawara, D Hashimoto, S Kimura, E Hayase, T Ara, S Takahashi, H Ohigashi, K Yoshioka, T Tateno, E Yokoyama, K Ebata, T Kondo, J Sugita, M Onozawa, T Iwanaga, T Teshima Intestinal Lymphatic Endothelial Cells Produce R-Spondin3 Sci Rep, 2018-07-16;8(1):10719. 2018-07-16 [PMID: 30013036] (Western Blot)
• G Greicius, Z Kabiri, K Sigmundsso, C Liang, R Bunte, MK Singh, DM Virshup PDGFR? + pericryptal stromal cells are the critical source of Wnts and RSPO3 for murine intestinal stem cells in vivo Proc. Natl. Acad. Sci. U.S.A., 2018-03-20;0(0):. 2018-03-20 [PMID: 29559533] (Bioassay, Human)
• S Liu, Y Yin, R Yu, Y Li, W Zhang R-spondin3-LGR4 signaling protects hepatocytes against DMOG-induced hypoxia/reoxygenation injury through activating ?-catenin Biochem. Biophys. Res. Commun., 2018-03-19;499(1):59-65. 2018-03-19 [PMID: 29555474] (Bioassay, Mouse)
• Y Yu, AH Moberly, JP Bhattarai, C Duan, Q Zheng, F Li, H Huang, W Olson, W Luo, T Wen, H Yu, M Ma The Stem Cell Marker Lgr5 Defines a Subset of Post-Mitotic Neurons in the Olfactory Bulb J. Neurosci., 2017-08-28;0(0):. 2017-08-28 [PMID: 28847812] (Bioassay, Mouse)
• X Yin, H Yi, L Wang, W Wu, X Wu, L Yu RSPOs facilitated HSC activation and promoted hepatic fibrogenesis Oncotarget, 2016-09-27;7(39):63767-63778. 2016-09-27 [PMID: 27572318] (Bioassay, Mouse)
• Cambier L, Plate M, Sucov H, Pashmforoush M Nkx2-5 regulates cardiac growth through modulation of Wnt signaling by R-spondin3. Development, 2014-08-01;141(15):2959-71. 2014-08-01 [PMID: 25053429] (Bioassay, Mouse)
• Li , Ji-Yao, Chai , Biaoxin, Zhang , Weizhen, Fritze , Danielle, Zhang , Chao, Mulholland , Michael LGR4 and its ligands, R-spondin 1 and R-spondin 3, regulate food intake in the hypothalamus of male rats. Endocrinology, 2013-11-26;155(2):429-40. 2013-11-26 [PMID: 24280058] (In Vivo, Mouse)
• Luo W, Rodriguez M, Valdez J, Zhu X, Tan K, Li D, Siwko S, Xin L, Liu M Lgr4 is a key regulator of prostate development and prostate stem cell differentiation. Stem Cells, 2013-11-01;31(11):2492-505. 2013-11-01 [PMID: 23897697] (Bioassay, Mouse)

Bioinformatics

• Gene Symbol: Rspo3
• Uniprot:
  • Q2TJ95()