Recombinant Mouse R-Spondin 1 Fc Chimera Protein, CF

Recombinant Mouse R-Spondin 1 Fc Chimera Protein (Catalog # 11566-RS) activates TCF reporter activity in HEK293 human embryonic kidney cells in the presence of Wnt-3a. The ED50 for this effect is 5.00-75.0 ng/mL.

2 μg/lane of Recombinant Mouse R‑Spondin 1 Fc Chimera Protein (Catalog # 11566-RS) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at SDS-PAGE 57-63 kDa and 110‑130 kDa, respectively.

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

Recombinant Mouse R-Spondin 1 Fc Chimera 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 5.00-75.0 ng/mL.
• Source: Chinese Hamster Ovary cell line, CHO-derived mouse R-Spondin 1 protein
  

  Mouse RSPO1 (Ser21-Pro207) Accession # NP_619624.2	IEGRMDP	Mouse IgG2a (Glu98-Lys330)
  N-terminus		C-terminus

• N-terminal Sequence: Ser21
• Structure / Form: Disulfide-linked homodimer
• Protein/Peptide Type: Recombinant Proteins
• 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: 57-63 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.
• Reconstitution Instructions: Reconstitute the 20 μg size at 200 μg/mL in PBS. Reconstitute all other sizes 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 1 Fc Chimera Protein, CF

• Cristin 3
• CRISTIN3
• FLJ40906Roof plate-specific spondin-1
• HRspo1
• roof plate-specific spondin
• RSPO
• RSPO1
• RSpondin 1
• R-Spondin 1
• R-spondin homolog (Xenopus laevis)
• RSPONDIN
• R-spondin1
• R-spondin-1

Background

R-Spondin 1 (RSPO1, Roof plate-specific Spondin 1), also known as cysteine-rich and single thrombospondin domain containing protein 3 (Cristin 3), is a 27 kDa secreted protein that shares ~40% amino acid (aa) identity with three other R-Spondin family members (1, 2). All R-Spondins regulate Wnt/ beta-Catenin signaling but have distinct expression patterns (1-3). R-Spondin 1 competes with the Wnt antagonist DKK-1 for binding to the Wnt co-receptors, Kremen and LRP-6, reducing their DKK-1-mediated internalization (4). However, reports are mixed on whether R-Spondin 1 binds LRP-6 directly (4-6). R-Spondin 1 is expressed in early development at the roof plate boundary and is thought to contribute to dorsal neural tube development (3, 7). In humans, rare disruptions of the R-Spondin 1 gene are associated with tendencies for XX sex reversal (phenotypic male) or hermaphroditism, indicating a role for R-Spondin 1 in gender-specific differentiation (7, 8). Mutations in R-Spondin 1 are also linked with palmoplantar keratoderma, abnormal thickening of the skin on the palms of the hands and soles of the feet (7, 8). Postnatally, R-Spondin 1 is expressed by neuroendocrine cells in the intestine, adrenal gland and pancreas, and by epithelia in kidney and prostate (9). Injection of recombinant R-Spondin 1 in mice causes activation of beta-catenin and proliferation of intestinal crypt epithelial cells, and ameliorates experimental colitis (9, 10). Interest in R-Spondin 1 as a cell culture supplement has grown with the expansion of the organoid field. R-Spondin 1 is widely used in organoid cell culture workflows as a vital component that promotes both growth and survival of 3D organoids (11).

Structurally similar to other R-Spondins, R-Spondin 1 contains two adjacent cysteine-rich furin-like domains (aa 34-135) with one potential N-glycosylation site, followed by a thrombospondin (TSP-1) motif (aa 147-207) and a region rich in basic residues (aa 211-263). Only the furin-like domains are needed for beta-catenin stabilization (2, 12). A putative nuclear localization signal at the C-terminus may allow some expression in the nucleus (13). Mouse R‑Spondin 1 shares 98%, 94%, 94%, 93%, 92% and 88% aa identity with rat, human, horse, cow, goat and dog RSPO-1, respectively, within aa 21‑209.

1. Chen, J.-Z. et al. (2002) Mol. Biol. Rep. 29:287.
2. Kim, K.-A. et al. (2006) Cell Cycle 5:23.
3. Nam, J.-S. et al. (2007) Gene Expr. Patterns 7:306.
4. Binnerts, M.E. et al. (2007) Proc. Natl. Acad. Sci. USA 104:14700.
5. Nam, J.-S. et al. (2006) J. Biol. Chem. 281:13247.
6. Wei, Q. et al. (2007) J. Biol. Chem. 282:15903.
7. Kamata, T. et al. (2004) Biochim. Biophys. Acta 1676:51.
8. Parma, P. et al. (2006) Nat. Genet. 38:1304.
9. Kim, K.-A. et al. (2005) Science 309:1256.
10. Zhao, J. et al. (2007) Gastroenterology 132:1331.
11. Drost and Clevers. (2018) Nature Reviews Cancer 18:407.
12. Kazanskaya, O. et al. (2004) Dev. Cell 7:525.
13. Tomaselli, S. et al. (2008) Hum. Mutat. 29:220.