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Recombinant Human R-Spondin 3 Protein

Catalog #: 3500-RS
17 Citations Datasheet / COA / SDS
Analyzed by SEC-MALS

Carrier Free

Catalog # Availability Size / Price Qty
3500-RS-025/CF

With Carrier

Catalog # Availability Size / Price Qty
3500-RS-025
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Recombinant Human R‑Spondin 3 Protein SEC-MALS.
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Product Details
Citations (17)
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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human R-Spondin 3 Protein Summary

Product Specifications

Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its ability to induce Topflash reporter activity in HEK293T human embryonic kidney cells. The ED50 for this effect is 0.500‑2.00 ng/mL in the presence of 5 ng/mL Recombinant Mouse Wnt‑3a (Catalog # 1324-WN).
Source
Chinese Hamster Ovary cell line, CHO-derived human R-Spondin 3 protein
Gln22-His272
Accession #
Q9BXY4
N-terminal Sequence
Analysis
No results obtained: Gln22 predicted
Predicted Molecular Mass
28.3 kDa
SDS-PAGE
36-45 kDa, reducing conditions

Product Datasheets

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3500-RS (with carrier)

Product Datasheet
COA
SDS

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3500-RS/CF (carrier free)

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.

3500-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% of human or bovine serun 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.

3500-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.

Scientific Data

SEC-MALS View Larger

Recombinant human R-Spondin 3 (Catalog # 3500-RS) has a molecular weight (MW) of 36.1 kDa as analyzed by SEC-MALS, suggesting that this protein is a monomer.  MW may differ from predicted MW due to post-translational modifications (PTMs) present (i.e. Glycosylation).

Bioactivity Recombinant Human R-Spondin 3 Protein Bioactivity View Larger

Recombinant Human R-Spondin 3 (Catalog # 3500-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. The ED50 for this effect is 0.50-2.00 ng/mL.

Bioactivity View Larger

The lot-to-lot consistency of Recombinant Human R-Spondin 3 (Catalog # 3500-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 3 from a different manufacturing run. The ED50 for this effect is 0.500-2.00 ng/mL in the presence of 5 ng/mL Recombinant Mouse Wnt-3a.

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 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
  1. Chen, J-Z. et al. (2002) Mol. Biol. Rep. 29:287.
  2. Kim, K.-A. et al. (2008) Mol. Biol. Cell 19:2588.
  3. Hendrickx, M. and L. Leyns (2008) Develop. Growth Differ. 50:229.
  4. Nam, J.-S. et al. (2007) Gene Expr. Patterns 7:306.
  5. Entrez Accession # EAW48114 and EAW48116.
  6. Kazanskaya, O. et al. (2008) Development 135:3655.
  7. Aoki, M. et al. (2007) Dev. Biol. 301:218.
  8. Binnerts, M.E. et al. (2007) Proc. Natl. Acad. Sci. USA 104:14700.
  9. Nam, J.-S. et al. (2006) J. Biol. Chem. 281:13247.
  10. Wei, Q. et al. (2007) J. Biol. Chem. 282:15903.
  11. 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

Citations for Recombinant Human R-Spondin 3 Protein

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

17 Citations: Showing 1 - 10
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  1. R-spondin 3 deletion induces Erk phosphorylation to enhance Wnt signaling and promote bone formation in the appendicular skeleton
    Authors: K Nagano, K Yamana, H Saito, R Kiviranta, AC Pedroni, D Raval, C Niehrs, F Gori, R Baron
    Elife, 2022-11-02;11(0):.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay
  2. The oncogenic transcription factor FOXQ1 is a differential regulator of Wnt target genes
    Authors: G Pizzolato, L Moparthi, S Söderholm, C Cantù, S Koch
    Journal of Cell Science, 2022-10-10;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Cell Culture
  3. Long-term culture, genetic manipulation and xenotransplantation of human normal and breast cancer organoids
    Authors: JF Dekkers, EJ van Vliet, N Sachs, JM Rosenbluth, O Kopper, HG Rebel, EJ Wehrens, C Piani, JE Visvader, CS Verissimo, SF Boj, JS Brugge, H Clevers, AC Rios
    Nature Protocols, 2021-03-10;0(0):.
    Species: Human
    Sample Types: Organoid
    Applications: Bioassay
  4. Development of a human primary gut-on-a-chip to model inflammatory processes
    Authors: C Beaurivage, A Kanapeckai, C Loomans, KS Erdmann, J Stallen, RAJ Janssen
    Scientific Reports, 2020-12-08;10(1):21475.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  5. RSPO3 impacts body fat distribution and regulates adipose cell biology in vitro
    Authors: NY Loh, JEN Minchin, KE Pinnick, M Verma, M Todor?evi?, N Denton, JE Moustafa, JP Kemp, CL Gregson, DM Evans, MJ Neville, KS Small, MI McCarthy, A Mahajan, JF Rawls, F Karpe, C Christodou
    Nat Commun, 2020-06-03;11(1):2797.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  6. Breast cancer organoids from a patient with giant papillary carcinoma as a high-fidelity model
    Authors: X Li, B Pan, X Song, N Li, D Zhao, M Li, Z Zhao
    Cancer Cell Int., 2020-03-18;20(0):86.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  7. Targeting the Wnt signaling pathway through R-spondin 3 identifies an anti-fibrosis treatment strategy for multiple organs
    Authors: M Zhang, M Haughey, NY Wang, K Blease, AM Kapoun, S Couto, I Belka, T Hoey, M Groza, J Hartke, B Bennett, J Cain, A Gurney, B Benish, P Castiglion, C Drew, J Lachowicz, L Carayannop, SD Nathan, J Distler, DA Brenner, K Hariharan, H Cho, W Xie
    PLoS ONE, 2020-03-11;15(3):e0229445.
    Species: Human
    Sample Types: Whole Cells
    Applications: IHC Control
  8. A cancer rainbow mouse for visualizing the functional genomics of oncogenic clonal expansion
    Authors: PG Boone, LK Rochelle, JD Ginzel, V Lubkov, WL Roberts, PJ Nicholls, C Bock, ML Flowers, RJ von Furste, BR Stripp, P Agarwal, AD Borowsky, RD Cardiff, LS Barak, MG Caron, HK Lyerly, JC Snyder
    Nat Commun, 2019-12-02;10(1):5490.
    Species: Human
    Sample Types: Whole Cells
    Applications: Cell Culture
  9. Differential activities and mechanisms of the four R-Spondins in potentiating Wnt/?-catenin signaling
    Authors: S Park, J Cui, WA Yu, L Wu, K Carmon, QJ Liu
    J. Biol. Chem., 2018-05-11;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  10. R-spondins can potentiate WNT signaling without LGRs
    Authors: AM Lebensohn, R Rohatgi
    Elife, 2018-02-06;7(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  11. Exercise-Induced Skeletal Muscle Adaptations Alter the Activity of Adipose Progenitor Cells
    Authors: D Zeve, DP Millay, J Seo, JM Graff
    PLoS ONE, 2016-03-25;11(3):e0152129.
    Species: Primate - Chlorocebus pygerythrus (Vervet Monkey)
    Sample Types: Whole Cells
    Applications: Bioassay
  12. Therapeutic Targeting of Tumor-Derived R-Spondin Attenuates beta-Catenin Signaling and Tumorigenesis in Multiple Cancer Types.
    Authors: Chartier C, Raval J, Axelrod F, Bond C, Cain J, Dee-Hoskins C, Ma S, Fischer M, Shah J, Wei J, Ji M, Lam A, Stroud M, Yen W, Yeung P, Cancilla B, O'Young G, Wang M, Kapoun A, Lewicki J, Hoey T, Gurney A
    Cancer Res, 2015-12-30;76(3):713-23.
    Species: Mouse
    Sample Types: In Vivo
    Applications: Bioassay
  13. RSPO-LGR4 functions via IQGAP1 to potentiate Wnt signaling.
    Authors: Carmon K, Gong X, Yi J, Thomas A, Liu Q
    Proc Natl Acad Sci U S A, 2014-03-17;111(13):E1221-9.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  14. RSPO2-LGR5 signaling has tumour-suppressive activity in colorectal cancer.
    Authors: Wu C, Qiu S, Lu L, Zou J, Li W, Wang O, Zhao H, Wang H, Tang J, Chen L, Xu T, Sun Z, Liao W, Luo G, Lu X
    Nat Commun, 2014-01-01;5(0):3149.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  15. Interaction with both ZNRF3 and LGR4 is required for the signalling activity of R-spondin.
    Authors: Xie, Yang, Zamponi, Raffaell, Charlat, Olga, Ramones, Melissa, Swalley, Susanne, Jiang, Xiaomo, Rivera, Daniel, Tschantz, William, Lu, Bo, Quinn, Lisa, Dimitri, Chris, Parker, Jefferso, Jeffery, Doug, Wilcox, Sheri K, Watrobka, Mike, LeMotte, Peter, Granda, Brian, Porter, Jeffrey, Myer, Vic E, Loew, Andreas, Cong, Feng
    EMBO Rep, 2013-10-29;14(12):1120-6.
    Species: Human
    Sample Types: Whole Cells
    Applications: Cell Culture
  16. Widespread potential for growth-factor-driven resistance to anticancer kinase inhibitors.
    Authors: Wilson TR, Fridlyand J, Yan Y, Penuel E, Burton L, Chan E, Peng J, Lin E, Wang Y, Sosman J, Ribas A, Li J, Moffat J, Sutherlin DP, Koeppen H, Merchant M, Neve R, Settleman J
    Nature, 2012-07-26;487(7408):505-9.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  17. ZNRF3 promotes Wnt receptor turnover in an R-spondin-sensitive manner.
    Authors: Hao HX, Xie Y, Zhang Y, Charlat O, Oster E, Avello M, Lei H, Mickanin C, Liu D, Ruffner H, Mao X, Ma Q, Zamponi R, Bouwmeester T, Finan PM, Kirschner MW, Porter JA, Serluca FC, Cong F
    Nature, 2012-04-29;485(7397):195-200.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay

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