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View LargerRecombinant Human R-Spondin 1 (Catalog # 4645-RS), in the presence of Recombinant Mouse Wnt-3a (Catalog # 1324-WN); 5 ng/mL), induces activation of beta-catenin in HEK293T cells measured using the Topflash assay (blue). The activity is approximately 7-fold greater than the competitor's R-Spondin 1 (red).
View LargerThe lot-to-lot consistency of Recombinant Human R-Spondin 1 (Catalog # 4645-RS) was assessed by testing the ability of three independent lots of the protein to stimulate activation of beta-Catenin using a TOPflash beta-Catenin/TCF reporter assay in the HEK293T human kidney cell line, in the presence of 5 ng/mL Recombinant Mouse Wnt-3a (1324-WN). Each trace on the graph represents data obtained from Recombinant Human R-Spondin 1 from a different manufacturing run. The ED50 for this effect is 1.00-10.0 ng/mL in the presence of 5 ng/mL Recombinant Mouse Wnt-3a.
View Larger1 µg/lane of Recombinant Human R-Spondin 1 was resolved with SDS-PAGE and visualized by silver staining under reducing (R) conditions, showing a single band at 39 kDa.
View LargeriPSC-derived human intestinal organoids were cultured using Cultrex™ UltiMatrix RGF Basement Membrane Extract (BME001-05) and intestinal organoid culture medium, which includes Recombinant Human R-Spondin 1 (Catalog # 4645-RS), Recombinant Human EGF (236-EG), Recombinant Human Noggin (6057-NG), and Recombinant Human Wnt-3a (5036-WN), along with the other reagents listed in the intestinal organoid culture medium recipe in the human intestinal organoid culture protocol. (A) Human intestinal organoids were stained using a Rat Anti-Human/Mouse/Rat Vimentin Monoclonal Antibody (MAB2105; green) and a Goat Anti-Human/Mouse Desmin Antigen Affinity-purified Polyclonal Antibody (AF3844; red) to visualize myofibroblast cells and counterstained with DAPI (5748; blue). (B) Human intestinal organoids were stained using a Goat Anti-Human/Mouse E-Cadherin Antigen Affinity-purified Polyclonal Antibody (AF748; green) and a Mouse Anti-Human MUC2 Monoclonal Antibody (Novus Biologicals, Catalog # NBP2-44431; red) and counterstained with DAPI (5748; blue).
View LargerAdult stem cells isolated from human descending colon were embedded in Cultrex UltiMatrix RGF Basement Membrane Extract (BME001-05) and cultured for 30 days in intestinal organoid culture medium, which includes Recombinant Human R-Spondin 1 (Catalog # 4645-RS), Recombinant Human EGF (236-EG), Recombinant Human Noggin (6057-NG), and Recombinant Human Wnt-3a (5036-WN), along with the other reagents listed in the intestinal organoid culture medium recipe in the human intestinal organoid culture protocol. (A) Organoids were fixed and stained with a Mouse Anti-Human MUC2 Monoclonal Antibody (Novus Biologicals; Catalog # NBP2-44431; green) to visualize intestinal goblet cells and counterstained with a Goat Anti-Human/Mouse E-Cadherin Antigen Affinity-purified Polyclonal Antibody (AF748; red) and DAPI (5748; blue). The image shown was taken at 10x magnification. (B) Organoids were fixed and stained with a Mouse Anti-Human Chromogranin A Monoclonal Antibody (MAB90981; green) to visualize enteroendocrine cells and counterstained with a Goat Anti-Human/Mouse E-Cadherin Antigen Affinity-purified Polyclonal Antibody (AF748; red) and DAPI (5748; blue). The image shown was taken at 20x magnification.
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.
4645-RS
| Formulation | Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein. |
| Reconstitution | Reconstitute at 100 μg/mL in PBS containing at least 0.1% human or bovine serum albumin. |
| 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.
|
4645-RS/CF
| Formulation | Lyophilized from a 0.2 μm filtered solution in PBS. |
| Reconstitution | Reconstitute at 100 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.
|
Recombinant Human R-Spondin 1 Protein Summary
Product Specifications
Ser21-Ala263
Analysis
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). Potential isoforms of 200 and 236 aa have an alternate, shorter N-terminus or are missing aa 146-208, respectively (14). Over aa 21-263, human R-Spondin 1 shares 89%, 87%, 92%, 91%, 91% and 89% aa identity with mouse, rat, horse, dog, goat, and cow RSPO-1, respectively.
- 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.
- UniProt # Q2MKA7.
参考图片
1 µg/lane of Recombinant Human
R-Spondin 1 was resolved with SDS-PAGE and visualized by silver staining under reducing (R) conditions, showing a single band at 39 kDa.
Recombinant Human R-Spondin 1 (Catalog # 4645-RS), in the presence of Recombinant Mouse Wnt‑3a (Catalog # 1324-WN; 5 ng/mL), induces activation of beta-catenin in HEK293T cells measured using the Topflash assay (blue). The activity is approximately 7-fold greater than the competitor's R-Spondin 1 (red).
危险品化学品经营许可证(不带存储) 许可证编号：沪(杨)应急管危经许[2022]202944(QY) 
营业执照(三证合一)