Human R-Spondin 1 / RSPO1 (21-146) Protein, His Tag

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INTRODUCTION

QC & Assay Protocol

Citation & Background

Synonym
RSPO1,CRISTIN3,FLJ40906,RP11-566C13.1,R-spondin-1
Source
Human R-Spondin 1 (21-146), His Tag(RS6-H4220) is expressed from human 293 cells (HEK293). It contains AA Ser 21 - Ala 146 (Accession # AAI14967 ).
Predicted N-terminus: Ser 21
Request for sequence
Molecular Characterization
This protein carries a polyhistidine tag at the C-terminus
The protein has a calculated MW of 14.6 kDa. The protein migrates as 19-25 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.
Endotoxin
Less than 1.0 EU per μg by the LAL method.

Purity
>90% as determined by SDS-PAGE.
Formulation
Lyophilized from 0.22 μm filtered solution in PBS, pH7.4 with trehalose as protectant.
Contact us for customized product form or formulation.
Reconstitution
Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
Storage
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please avoid repeated freeze-thaw cycles.
This product is stable after storage at:
1. -20°C to -70°C for 12 months in lyophilized state;
2. -70°C for 3 months under sterile conditions after reconstitution.

SDS-PAGE

Human R-Spondin 1 (21-146), His Tag on SDS-PAGE under reducing (R) condition. The gel was stained with Coomassie Blue. The purity of the protein is greater than 90%.

Bioactivity-Organoid culture

Human EGF (Cat. EGF-H52H3), Noggin (Cat. NON-H5257) and R-spondin1 (Cat. RS6-H4220) could support the growth of mouse intestinal organoids.

Bioactivity-ELISA

Immobilized Human R-Spondin 1 (21-146), His Tag (Cat. No. RS6-H4220) at 5 μg/mL (100 μL/well) can bind Human RNF43, Fc Tag (Cat. No. RN3-H5256) with a linear range of 0.01-0.156 μg/mL (QC tested).

Protocol

Immobilized Human R-Spondin 1 (21-146), His Tag (Cat. No. RS6-H4220) at 5 μg/mL (100 μL/well) can bind Human LRP-6 (20-630), Mouse IgG2a Fc Tag (Cat. No. LR6-H5253) with a linear range of 0.039-2.5 μg/mL (Routinely tested).

Protocol

Background
R-spondin-1 is also known as Roof plate-specific Spondin 1 (RSPO1) and cysteinerich and single thrombospondin domain containing protein 3 (Cristin 3), which is a secreted protein which belongs to the R-Spondin family and encodes a secreted activator protein with two cystein-rich, furin-like domains and one thrombospondin type 1 domain. All Rspondins regulate Wnt/β-catenin signaling, but have distinct expression patterns. Like other R-Spondins, R-Spondin-1 contains two adjacent cysteinerich furinlike domains (aa 34-135) with one potential N-glycosylation site, followed by a thrombospondin (TSP1) motif (aa 147-207) and a region rich in basic residues (aa 211-263). Only the furinlike domains are needed for β-catenin stabilization. A putative nuclear localization signal at the C-terminus may allow some expression in the nucleus.
Clinical and Translational Updates
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