Recombinant Mouse R-Spondin 3 mFc Protein, CF New

Catalog #: 11540-RS Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
11540-RS-010
11540-RS-500

Recombinant Mouse R‑Spondin 3 mFc Protein Bioactivity.

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Product Details

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

Recombinant Mouse R-Spondin 3 mFc 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 ability to activate TCF reporter activity in HEK293 human embryonic kidney cells in the presence of Wnt-3a. The ED₅₀ for this effect is 0.300-3.60 ng/mL.

Source

Chinese Hamster Ovary cell line, CHO-derived mouse R-Spondin 3 protein

Mouse RSPO03 (Gln22, Met33-Gly209) Accession # Q2TJ95.2	IEGRMD	Mouse IgG_2a (Glu98-Lys330)
N-terminus		C-terminus

Accession #

Q2TJ95.2

N-terminal Sequence
Analysis

Gln22 (blocked) & Met 33

Structure / Form

Disulfide-linked homodimer

Predicted Molecular Mass

48 kDa

SDS-PAGE

54-65 kDa, under reducing conditions

Product Datasheets

11540-RS

Product Datasheet

COA

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.

11540-RS

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

Bioactivity

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Recombinant Mouse R-Spondin 3 mFc Protein (Catalog # 11540-RS) activates TCF reporter activity in HEK293 human embryonic kidney cells in the presence of Wnt-3a. The ED₅₀ for this effect is 0.300-3.60 ng/mL.

SDS-PAGE

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2 μg/lane of Recombinant Mouse R‑Spondin 3 mFc Protein (Catalog # 11540-RS) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 54-65 kDa and 110-130 kDa, respectively.

Reconstitution Calculator

Background: R-Spondin 3

R-Spondin 3 (RSPO3, roof plate-specific spondin 3), also called cysteine-rich and single thrombospondin domain containing-1 (Cristin 1), is an ~31 kDa secreted protein that shares ~40% aa identity with the other three R‑Spondin family members (1, 2). All are positive modulators of Wnt/ beta -catenin signaling, but each has a distinct expression pattern (1-4). Like other R‑spondins, R‑Spondin 3 contains two adjacent cysteine-rich furin‑like domains (amino acids (aa) 35-135) with one potential N‑glycosylation site (aa 36), followed by a thrombospondin (TSP-1) motif (aa 147-207) and a region rich in basic residues (aa 211-269). Only the furin-like domains are needed for beta -catenin stabilization (2). Within aa 21-209, mouse R‑Spondin 3 shares 93%, 97%, 96%, 95% and 91% aa identity with human, rat, equine, bovine and canine R‑Spondin 3, respectively. Potential isoforms of 217, 224 and 252 aa are divergent or truncated at the C terminus; the 252 aa form also lacks aa 4‑33 at the N‑terminus (5). Mouse R‑Spondin 3 is critical for development of the placental labyrinthine layer, probably by promoting VEGF expression and thus vascular development (6, 7). It is also essential for expression of the placenta-specific transcription factor, Gcm1. In the mouse embryo, R‑Spondin 3 is often expressed by or located near endothelial cells (6). It is found in the roof plate, tail, somites, otic vesicles, cephalic mesoderm, truncus arteriosus, atrioventricular canal of the developing heart, and strongly but transiently in developing limbs (4, 7). R‑Spondins regulate Wnt/ beta -catenin by competing with the Wnt antagonist DKK-1 for binding to the Wnt co-receptors LRP-6 and Kremen, reducing their DKK‑1‑mediated internalization (8, 9). Reports differ on whether R‑Spondins bind LRP-6 directly (8-10). R‑Spondin 3 has also been identified as an oncogene (11).

References

Chen, J-Z. et al. (2002) Mol. Biol. Rep. 29:287.
Kim, K.-A. et al. (2008) Mol. Biol. Cell 19:2588.
Hendrickx, M. and L. Leyns (2008) Develop. Growth Differ. 50:229.
Nam, J.-S. et al. (2007) Gene Expr. Patterns 7:306.
Entrez Accession # BAB28811, BAC36296 and EDL04842.
Kazanskaya, O. et al. (2008) Development 135:3655.
Aoki, M. et al. (2007) Dev. Biol. 301:218.
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.
Theodorou, V. et al. (2007) Nat. Genet. 6:759.

Long Name

Roof Plate-specific Spondin 3

Entrez Gene IDs

84870 (Human); 72780 (Mouse); 102132009 (Cynomolgus Monkey)

Alternate Names

Cristin 1; CRISTIN1; FLJ14440; hPWTSR; hRspo3; Protein with TSP type-1 repeat; PWTSR; Roof plate-specific spondin-3; RSPO3; R-spondin 3 homolog (Xenopus laevis); RSpondin 3; R-Spondin 3; R-spondin-3; Thrombospondin type-1 domain-containing protein 2; thrombospondin, type I, domain containing 2; THSD2