Recombinant Human R-Spondin 1 Biotinylated Protein, CF

Biotinylated
Catalog # Availability Size / Price Qty
BT4645B-025
Biotinylated Recombinant Human R‑Spondin 1 Protein Binding Activity.
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Recombinant Human R-Spondin 1 Biotinylated Protein, CF Summary

Product Specifications

Purity
>95%, 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.
Activity
Measured by its binding ability in a functional ELISA. When Recombinant Human Lgr5/GPR49 Fc Chimera (Catalog # 8078-GP) is immobilized at 2 µg/mL (100 µL/well), Biotinylated Recombinant Human R-Spondin 1 (Catalog # BT4645B) binds with an ED50 of 4.50-54.0 ng/mL.
Source
Chinese Hamster Ovary cell line, CHO-derived human R-Spondin 1 protein
Ser21-Ala263
Accession #
N-terminal Sequence
Analysis
Ser21 & Arg31
Structure / Form
Biotinylated via amines
Predicted Molecular Mass
25.6 kDa
SDS-PAGE
29-42 kDa, under reducing conditions.

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BT4645B

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.

BT4645B

Formulation Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution Reconstitute at 250 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.

Scientific Data

Binding Activity View Larger

When Recombinant Human Lgr5/GPR49 Fc Chimera (8078-GP) is immobilized at 2 µg/mL (100 µL/well), Biotinylated Recombinant Human R-Spondin 1 Protein (Catalog # BT4645B) binds with an ED50 of 4.50-54.0 ng/mL.

SDS-PAGE View Larger

2 μg/lane of Biotinylated Recombinant Human R‑Spondin 1 Protein (Catalog # BT4645B) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 29‑42 kDa.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: R-Spondin 1

R-Spondin 1 (RSPO1), also known as Cristin 3, is a 27 kDa secreted protein in the R-Spondin family of Wnt/ beta -catenin signaling regulators (1). These proteins contain two adjacent cysteine-rich furin-like domains followed by a thrombospondin (TSP-1) motif and a region rich in basic residues. Mature human R-Spondin 1 shares 87% amino acid sequence identity with mouse and rat R-Spondin 1 (2). Alternative splicing generates additional isoforms that have a substituted N-terminus or lack the TSP-1 domain. R-Spondin 1 enhances canonical Wnt/ beta -catenin signaling by competing with the Wnt antagonist Dkk-1, binding to Frizzled-8, Kremen, LRP-6, Lgr4, Lgr5, and Lgr6, and enhancing cell surface availability of Wnt receptors (3-11). R-Spondin 1 functions in dorsal neural tube development (12) as well as male and female germ cell development (7, 10, 13). It also induces bone formation (6), intestinal crypt cell proliferation (14), angiogenesis (7, 15), and insulin secretion from pancreatic beta cells (11). 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 (16). Over the last several years, the understanding of the regulatory mechanisms and functional roles of RSPOs in many biological contexts has increased. Particularly, because a leucine-rich repeat containing G protein-coupled receptor 5 (LGR5), a stem cell marker originally identified as a marker for intestinal stem cells, and two closely related proteins, LGR4 and LGR6, were identified as cognate receptors for RSPOs, significant research progress has been made in understanding the functional roles of RSPO/LGR signaling in stem cell biology (17).

References
  1. Jin, Y.R. and J.K. Yoon (2012) Int. J. Biochem. Cell Biol. 44:2278.
  2. Chen, J-Z. et al. (2002) Mol. Biol. Rep. 29:287.
  3. Binnerts, M.E. et al. (2007) Proc. Natl. Acad. Sci. USA 104:14700.
  4. Nam, J.-S. et al. (2006) J. Biol. Chem. 281:13247.
  5. Wei, Q. et al. (2007) J. Biol. Chem. 282:15903.
  6. Kronke, G. et al. (2010) Arthritis Rheum. 62:2303.
  7. Caruso, M. et al. (2015) PLoS One 10:e0124213.
  8. Hao, H.-X. et al. (2012) Nature 485:195.
  9. de Lau, W. et al. (2011) Nature 476:293.
  10. Chassot, A.-A. et al. (2011) PLoS One 6:e25641.
  11. Wong, V.S.C et al. (2010) J. Biol. Chem. 285:21292.
  12. Kamata, T. et al. (2004) Biochim. Biophys. Acta 1676:51.
  13. Chassost, A.-A. et al. (2012) Development 139:4461.
  14. Kim, K.-A. et al. (2005) Science 309:1256.
  15. Gore, A.V. et al. (2011) Development 138:4875.
  16. Drost and Clevers. (2018) Nature Reviews Cancer 18:407.
  17. Raslan, H. et al. (2019) J. Biochem. Biocell. 106:26-34.
Long Name
Roof Plate-specific Spondin 1
Entrez Gene IDs
284654 (Human); 192199 (Mouse); 102122369 (Cynomolgus Monkey)
Alternate Names
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

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