Recombinant Mouse R-Spondin 1 Fc Chimera Protein, CF New
Recombinant Mouse R-Spondin 1 Fc Chimera Protein, CF Summary
Product Specifications
Mouse RSPO1 (Ser21-Pro207) Accession # NP_619624.2 | IEGRMDP | Mouse IgG2a (Glu98-Lys330) |
N-terminus | C-terminus | |
Analysis
Product Datasheets
Carrier Free
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
11566-RS
Formulation | Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. |
Reconstitution | Reconstitute the 20 μg size at 200 μg/mL in PBS. Reconstitute all other sizes at 500 μg/mL in PBS. |
Shipping | The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. |
Stability & Storage: | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
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Scientific Data
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.
Reconstitution Calculator
Background: R-Spondin 1
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.
- Chen, J.-Z. et al. (2002) Mol. Biol. Rep. 29:287.
- Kim, K.-A. et al. (2006) Cell Cycle 5:23.
- Nam, J.-S. et al. (2007) Gene Expr. Patterns 7:306.
- Binnerts, M.E. et al. (2007) Proc. Natl. Acad. Sci. USA 104:14700.
- Nam, J.-S. et al. (2006) J. Biol. Chem. 281:13247.
- Wei, Q. et al. (2007) J. Biol. Chem. 282:15903.
- Kamata, T. et al. (2004) Biochim. Biophys. Acta 1676:51.
- Parma, P. et al. (2006) Nat. Genet. 38:1304.
- Kim, K.-A. et al. (2005) Science 309:1256.
- Zhao, J. et al. (2007) Gastroenterology 132:1331.
- Drost and Clevers. (2018) Nature Reviews Cancer 18:407.
- Kazanskaya, O. et al. (2004) Dev. Cell 7:525.
- Tomaselli, S. et al. (2008) Hum. Mutat. 29:220.
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