Recombinant Mouse R-Spondin 3 mFc Protein, CF

Recombinant Mouse R-Spondin 3 mFc Protein (Catalog # 11540-RS) activates TCF reporter activity in HEK293 human embryonic kidney cells in the presence of Wnt-3a. The ED50 for this effect is 0.300-3.60 ng/mL.

2 μg/lane of Recombinant Mouse R‑Spondin 3 mFc Protein (Catalog # 11540-RS) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 54-65 kDa and 110-130 kDa, respectively.

• Reactivity: Mu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Mouse R-Spondin 3 mFc Protein, CF Summary

• Details of Functionality: Measured by its ability to activate TCF reporter activity in HEK293 human embryonic kidney cells in the presence of Wnt-3a. The ED₅₀ for this effect is 0.300-3.60 ng/mL.
• Source: Chinese Hamster Ovary cell line, CHO-derived mouse R-Spondin 3 protein
  

  Mouse RSPO03 (Gln22, Met33-Gly209) Accession # Q2TJ95.2	IEGRMD	Mouse IgG2a (Glu98-Lys330)
  N-terminus		C-terminus

• Accession #: Q2TJ95.2
• N-terminal Sequence: Gln22 (blocked) & Met 33
• Structure / Form: Disulfide-linked homodimer
• Protein/Peptide Type: Recombinant Proteins
• 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: 48 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: 54-65 kDa, under 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 Trehalose.
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Reconstitution Instructions: Reconstitute at 500 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 mFc Protein, CF

• 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.

Bioinformatics

• Uniprot:
  • Q2TJ95.2()