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Human R-Spondin 3 Antibody

R&D Systems, part of Bio-Techne | Catalog # AF35001

R&D Systems, part of Bio-Techne
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AF35001
AF35001-SP

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Neutralization

Cited:

Neutralization

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG

Product Specifications

Immunogen

Chinese hamster ovary cell line CHO-derived recombinant human R-Spondin 3
Gln22-His272
Accession # Q9BXY4

Specificity

Detects human R-Spondin 3 in direct ELISAs. In direct ELISAs, less than 10% cross-reactivity with recombinant mouse R-Spondin 3 is observed.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Endotoxin Level

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

Applications for Human R-Spondin 3 Antibody

Application
Recommended Usage

Neutralization

Measured by its ability to neutralize R‑Spondin 3-induced activation of beta‑Catenin response in the HEK293T human embryonic kidney cell line in a Topflash Luciferase assay. The Neutralization Dose (ND50) is typically 0.3-1.2 µg/mL in the presence of 10 ng/mL Recombinant Human R‑Spondin 3 and 5 ng/mL Recombinant Mouse Wnt‑3a.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL. For liquid material, refer to CoA for concentration.

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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store 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.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

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

Alternate Names

Cristin 1, PWTSR, RSPO3, RSpondin 3, THSD2

Entrez Gene IDs

84870 (Human); 72780 (Mouse); 102132009 (Cynomolgus Monkey)

Gene Symbol

RSPO3

UniProt

Additional R-Spondin 3 Products

Product Documents for Human R-Spondin 3 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human R-Spondin 3 Antibody

For research use only

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