Recombinant Human R-spondin 3, Animal-Free Protein

Catalog #: Qk032 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
Qk032-0050
Qk032-0500

Recombinant Human R-spondin 3, Animal-Free Protein Bioactivity

2 Images

Product Details

Reviews

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

Recombinant Human R-spondin 3, Animal-Free Protein Summary

Product Specifications

Endotoxin Level

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

Activity

No significant difference between EC₅₀ of reference and test lots

Source

E. coli-derived human R-Spondin 3 protein

Accession #

Q9BXY4.1

Predicted Molecular Mass

17 kDa

SDS-PAGE

Monomeric R-spondin 3 protein only

Product Datasheets

QK032

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.

Qk032

Formulation	Lyophilized from acetonitrile/TFA
Reconstitution	Resuspend in 10 mM HCl at >100 µg/ml, prepare single use aliquots, add carrier protein if desired.
Shipping	The product is shipped lyophilized at ambient temperture, on ice blocks or dry ice. Shipping at ambient temperture does not affect the bioactivity or stability of the protein. Upon reciept, store immediately at the conditions stated below.
Stability & Storage:	Store lyophilized protein between -20 and -80 °C until the date of expiry. Avoid freeze-thaw cycles.

Scientific Data

Bioactivity

View Larger

Cells transfected with reporter TOP-FLASH are treated in triplicate with increasing concentration of R-spondin 3 in the presence of Wnt-conditioned media (1:8 dilution). RSP1 enhances Wnt-beta catenin signaling in HEK239T cells. Cells are grown overnight, and luciferase activity is measured and normalized.EC₅₀ = 14 ng/ml (0.8 nM).

SDS-PAGE

View Larger

R-spondin 3 bioactive domain migrates at 17 kDa in non-reducing (-beta ME) conditions and upon reduction (+ beta ME).Purified recombinant protein (7 µg) was resolved using 15% w/v SDS-PAGE in reduced (+ beta -mercaptoethanol, R) and non-reduced conditions (NR) and stained with Coomassie Brilliant Blue R250.

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% amino acid (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 (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, human R-Spondin 3 shares 93%, 92%, 97%, 96% and 92% aa identity with mouse, rat, equine, bovine and canine R-Spondin 3, respectively. Potential isoforms of 279 and 297 aa diverge at aa 210 and 276, respectively (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 # EAW48114 and EAW48116.
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

Product Specific Notices

The above product was manufactured, tested and released by R&D System's contract manufacturer, Qkine Ltd, at 1 Murdoch House, Cambridge, UK, CB5 8HW. The product is for research use only and not for the diagnostic or theraputic use.