Human R-Spondin 4 DuoSet ELISA Summary
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
- Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
- Development protocols are provided to guide further assay optimization
- Assay can be customized to your specific needs
- Economical alternative to complete kits
- Capture Antibody
- Detection Antibody
- Recombinant Standard
- Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)
Other Reagents Required
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or equivalent
Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)
Stop Solution: 2 N H2SO4 (Catalog # DY994)
Microplates: R&D Systems (Catalog # DY990), or equivalent
Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent
*For the Reagent Diluent and Blocking Buffer recommended for a specific DuoSet ELISA Development Kit, please see the product
Human R-Spondin 4 ELISA Standard Curve
Preparation and Storage
Background: R-Spondin 4
R-Spondin 4 (RSPO4, roof plate-specific spondin 4), also called cysteine-rich and single thrombospondin domain containing-4 (Cristin 4), is an ~33 kDa secreted heparin-binding protein that shares ~35% amino acid (aa) identity with three other R-Spondin family members. All are positive modulators of Wnt/beta-Catenin signaling, but vary in activity. 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. Like other R-spondins, human R-Spondin 4 (228 aa) contains a signal sequence (aa 1-19), two adjacent cysteine-rich furin-like domains (aa 85-128) with one potential tyrosine phosphorylation site (aa 114), followed by a thrombospondin (TSP-1) motif (aa 137-197) and a region rich in basic residues (aa 199-228). Mature human R-Spondin 4 shares 81%, 81%, 84%, 84% and 86% aa identity with mouse, rat, equine, canine and bovine R-Spondin 4, respectively. Of two potential isoforms, one lacks the TSP-1 domain, while another terminates at aa 224. Each R-Spondin has a distinct expression pattern. In the mouse, R-Spondin 4 mRNA is found during development of limb bud mesenchyme, nail beds, heart and teeth. In humans, mutations of R-Spondin 4 have been found to cause anonychia, a condition in which fingernails and toenails are absent.
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