Recombinant Human R-Spondin 1 Biotinylated Protein, CF

Catalog #: BT4645B Datasheet / COA / SDS

Biotinylated

Catalog #	Availability	Size / Price	Qty
BT4645B-025

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

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 #

Q2MKA7.1

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.

Product Datasheets

BT4645B

Product Datasheet

COA

SDS

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

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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

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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

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

Jin, Y.R. and J.K. Yoon (2012) Int. J. Biochem. Cell Biol. 44:2278.
Chen, J-Z. et al. (2002) Mol. Biol. Rep. 29:287.
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.
Kronke, G. et al. (2010) Arthritis Rheum. 62:2303.
Caruso, M. et al. (2015) PLoS One 10:e0124213.
Hao, H.-X. et al. (2012) Nature 485:195.
de Lau, W. et al. (2011) Nature 476:293.
Chassot, A.-A. et al. (2011) PLoS One 6:e25641.
Wong, V.S.C et al. (2010) J. Biol. Chem. 285:21292.
Kamata, T. et al. (2004) Biochim. Biophys. Acta 1676:51.
Chassost, A.-A. et al. (2012) Development 139:4461.
Kim, K.-A. et al. (2005) Science 309:1256.
Gore, A.V. et al. (2011) Development 138:4875.
Drost and Clevers. (2018) Nature Reviews Cancer 18:407.
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