- Other Names
- Roof plate-specific spondin-2, R-spondin-2, CRISTIN2, RSPO-2, R-Spondin 2, RSPO2
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- Product Citations
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R-Spondin 2 (RSPO2) was initially discovered by an expression screen to identify modulators of Wnt signaling in Xenopus. The R-Spondin protein family consists of four members, R-Spondin 1 through R-Spondin 4. These proteins feature a hydrophobic signal peptide, two adjacent cystein-rich, furin-like domains and one thrombospondin type 1 domain. These are secreted proteins that are processed through a canonical secretory pathway. R-Spondin 2 and R-Spondin 3 bind heparin sulfate proteoglycans on the cell surface whereas R-Spondin 1 and R-Spondin 4 do not show significant interaction with heparin sulfate proteoglycans. All R-Spondin family members regulate the Wnt signaling pathway. RSPO2 enhances Wnt signaling through the inhibition of ubiquitin E3 ligases RNF43/ZNRF3. RSPO2 also activates the canonical Wnt signaling pathway by acting as a ligand for LGR4-6 receptors. Upon binding to LGR4-6 (LGR4, LGR5 or LGR6), LGR4-6 associate with phosphorylated LRP6 and frizzled receptors that are activated by extracellular Wnt receptors, triggering the canonical Wnt signaling pathway to increase expression of target genes. A chromosomal translocation including the RSPO2 locus that results in the formation of a gene fusion has been identified in multiple human cancers. Other diseases associated with RSPO2 include humerofemoral hypoplasia with radiotibial ray deficiency and tetra-amelia syndrome 2.Product Details
- Human RSPO2, amino acid Gln22-Gly205 (Accession # Q6UXX9), with a C-terminal hIgG1 Fc tag, was expressed in 293E cells.
- Molecular Mass
- The 423 amino acid recombinant protein has a predicted molecular mass of approximately 48.2 kD. The DTT-reduced protein migrates at approximately 55 kD and and non-reduced protein migrates at approximately 100 kD by SDS-PAGE. The predicted N-terminal amino acid is Gln.
- >95%, as determined by Coomassie stained SDS-PAGE.
- 0.22 µm filtered protein solution is in PBS, pH 7.2.
- Endotoxin Level
- Less than 0.01 ng per µg cytokine as determined by the LAL method.
- 10 and 25 µg sizes are bottled at 200 µg/mL. 100 µg and larger sizes are lot-specific and bottled at the concentration indicated on the vial (please contact technical support for concentration, or use our Lookup tool if you have a lot number.)
- Storage & Handling
- Unopened vial can be stored between 2°C and 8°C for up to 2 weeks, at -20°C for up to six months, or at -70°C or colder until the expiration date. For maximum results, quick spin vial prior to opening. The protein can be aliquoted and stored at -20°C or colder. Stock solutions can also be prepared at 50 - 100 µg/mL in appropriate sterile buffer, carrier protein such as 0.2 - 1% BSA or HSA can be added when preparing the stock solution. Aliquots can be stored between 2°C and 8°C for up to one week and stored at -20°C or colder for up to 3 months. Avoid repeated freeze/thaw cycles.
When human RSPO2-Fc Chimera is coated at 0.5 µg/mL (100 µL/well), biotinylated human RNF43-Fc Chimera binds with an EC50 of 2.5 - 10 ng/mL. HRP Avidin (Cat. No. 405103) was used to detect the binding.
Human RSPO2-Fc Chimera also induces Topflash reporter activity in HEK293T human embryonic kidney cells in the presence of 5 ng/mL mWnt3a (Cat. No. 772301) with an EC50 of 0.4 - 1.6 ng/mL.
- Application Notes
BioLegend carrier-free recombinant proteins provided in liquid format are shipped on blue ice. Our comparison testing data indicates that when handled and stored as recommended, the liquid format has equal or better stability and shelf-life compared to commercially available lyophilized proteins after reconstitution. Our liquid proteins are verified in-house to maintain activity after shipping on blue ice and are backed by our 100% satisfaction guarantee. If you have any concerns, contact us at email@example.com.
Broadly distributed in normal adult tissues including brain, endocrine tissues, muscle tissues, lung, pancreas, gastrointestinal tract, kidney, testis, and ovary
- An activator of the canonical Wnt signaling pathway that plays a crucial role in limb specification during the embryonic development.
- Intestinal epithelial cells
- LGR4-6, RNF43/ZNRF3
- Measured by its ability to bind RNF43 and induce Topflash reporter activity in HEK293T human embryonic kidney cells.
- Cell Type
- Embryonic Stem Cells, Mesenchymal cells, Mesenchymal Stem Cells, Neural Stem Cells
- Biology Area
- Cell Proliferation and Viability, Stem Cells
- Molecular Family
- Growth Factors
- Antigen References
- Kim KA, et al. 2005. Science. 309:1256-9
- Parma P, et al. 2006. Nature Genet. 38:1304-9
- Chassot A, et al. 2008. Hum Molec Genet. 17:1264-77
- Zhou WJ, et al. 2013. Nature. 501:107-11
- Gene ID
- 340419 View all products for this Gene ID
- View information about R-Spondin 2 on UniProt.org
- Does specific activity of a recombinant protein vary between lots?
Specific activity will vary for each lot and for the type of experiment that is done to validate it, but all passed lots will have activity within the established ED50 range for the product and we guarantee that our products will have lot-to-lot consistency. Please conduct an experiment-specific validation to find the optimal ED50 for your system.
- Have your recombinants been tested for stability?
Our testing shows that the recombinant proteins are able to withstand room temperature for a week without losing activity. In addition the recombinant proteins were also found to withstand four cycles of freeze and thaw without losing activity.
- How do you convert activity as an ED50 in ng/ml to a specific activity in Units/mg?
- Use formula Specific activity (Units/mg) = 10e6/ ED50 (ng/mL)
- How does the activity of your recombinant proteins compare to competitors?
We quality control each and every lot of recombinant protein. Not only do we check its bioactivity, but we also compare it against other commercially available recombinant proteins. We make sure each recombinant protein’s activity is at least as good as or better than the competition’s. In order to provide you with the best possible product, we ensure that our testing process is rigorous and thorough. If you’re curious and eager to make the switch to BioLegend recombinants, contact your sales representative today!
- What is the specific activity or ED50 of my recombinant protein?
The specific activity range of the protein is indicated on the product datasheets. Because the exact activity values on a per unit basis can largely fluctuate depending on a number of factors, including the nature of the assay, cell density, age of cells/passage number, culture media used, and end user technique, the specific activity is best defined as a range and we guarantee the specific activity of all our lots will be within the range indicated on the datasheet. Please note this only applies to recombinants labeled for use in bioassays. ELISA standard recombinant proteins are not recommended for bioassay usage as they are not tested for these applications.