Recombinant Human R-Spondin 1 Protein Best Seller

Catalog # 4645-RS | R&D Systems, Inc. a Bio-Techne Brand
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4645-RS-01M/CF
4645-RS-025/CF
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4645-RS-100/CF
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4645-RS-01M
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Key Product Details

Accession #

Q2MKA7

Source

CHO

Conjugate

Unconjugated

Applications

Bioactivity

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human R-Spondin 1 protein
Ser21-Ala263

Purity

>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Ser21 & Arg31

Predicted Molecular Mass

25.6 kDa

SDS-PAGE

39 kDa, reducing conditions

Activity

Measured by its ability to induce Topflash reporter activity in HEK293T human embryonic kidney cells.
The ED50 for this effect is 1.00-10.0 ng/mL in the presence of 5 ng/mL Recombinant Mouse Wnt‑3a (Catalog # 1324-WN).

Scientific Data Images for Recombinant Human R-Spondin 1 Protein

Recombinant Human R-Spondin 1 Protein Bioactivity

Recombinant Human R-Spondin 1 Protein Bioactivity

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).

Recombinant Human R-Spondin 1 Protein Lot-to-Lot Consistency

The 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.
Recombinant Human R-Spondin 1 Protein SDS-PAGE

Recombinant Human R-Spondin 1 Protein SDS-PAGE

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.
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Formulation, Preparation and Storage

Carrier Free
What does CF mean?

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.

What formulation is right for me?

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.

Carrier: 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.
  • 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.
Carrier Free: 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.
  • 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.

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.

References

  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.
  14. UniProt # Q2MKA7.

Long Name

Roof Plate-specific Spondin 1

Alternate Names

Cristin 3, CRISTIN3, HRspo1, RSPO, RSPO1, RSpondin 1

Entrez Gene IDs

284654 (Human); 192199 (Mouse); 102122369 (Cynomolgus Monkey)

Gene Symbol

RSPO1

UniProt

Q2MKA7

Product Documents for Recombinant Human R-Spondin 1 Protein

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS 4645-RS (with carrier)
Download SDS 4645-RS/CF (carrier free)

Product Specific Notices for Recombinant Human R-Spondin 1 Protein

For research use only

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Customer Reviews for Recombinant Human R-Spondin 1 Protein (6)

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  • Recombinant Human R-Spondin 1 Protein
    Name: Anonymous
    Application: 3D cell culture/organoids.
    Verified Customer | Posted 10/11/2023
    Recombinant Human R-Spondin 1 Protein 4645-RS
  • Recombinant Human R-Spondin 1 Protein
    Name: Andres Ocampo Carrillo
    Application: 3D culture of cells
    Verified Customer | Posted 03/04/2023
    R-Spondin was used for colonocyte isolation and culturing from Sprague-Dawley rats in 3D culture. Results showed a purified culture as spheroids-like, following a standard isolation protocol for these cells
    Recombinant Human R-Spondin 1 Protein 4645-RS
  • Recombinant Human R-Spondin 1 Protein
    Name: Anonymous
    Application: Stem/Immune cell maintenance or differentiation
    Verified Customer | Posted 09/26/2021
    Used for the differentiation of iPSC into intestinal organoids, at a concentration of 500ng/mL
  • Recombinant Human R-Spondin 1 Protein
    Name: Ermali Aliaj
    Application: Cell Culture
    Verified Customer | Posted 01/02/2021
  • Recombinant Human R-Spondin 1 Protein
    Name: Anonymous
    Application: Stem/Immune cell maintenance or differentiation
    Verified Customer | Posted 07/24/2020
  • Recombinant Human R-Spondin 1 Protein
    Name: Anonymous
    Application: In vitro bioactivity in cell culture
    Verified Customer | Posted 03/06/2020

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