Mouse R-Spondin 1 Antibody

R&D Systems | Catalog # AF3474

R&D Systems

Key Product Details

Species Reactivity

Validated:

Mouse

Cited:

Human, Mouse

Applications

Validated:

Immunohistochemistry, Western Blot

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Bioassay

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all R-Spondin 1 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant mouse R-Spondin 1 (R&D Systems, Catalog # 3474-RS)
Ser21-Gly209
Accession # Q9Z132

Specificity

Detects mouse R-Spondin 1 in direct ELISAs and Western blots. In direct ELISAs, approximately 60% cross‑reactivity with recombinant human R-Spondin 1 is observed and less than 5% cross-reactivity with recombinant mouse (rm) R-Spondin 3 and rmR-Spondin 4 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Mouse R-Spondin 1 Antibody

R-Spondin 1 antibody in Mouse Ovary by Immunohistochemistry (IHC-Fr).

R‑Spondin 1 in Mouse Ovary.

R-Spondin 1 was detected in immersion fixed frozen sections of E13.5 mouse ovary using Goat Anti-Mouse R-Spondin 1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3474) at 10 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.
Detection of Mouse R-Spondin 1 by Western Blot

Detection of Mouse R-Spondin 1 by Western Blot

RSPO1 protein content and distribution pattern during male gonad development.Left panel: (Upper part) Western blot analysis of RSPO1 in 13.5, 16.5 and 18.5 dpc embryonic male gonads, and 13.5 dpc male germ cells. As RSPO1 western blot negative control 17.5 dpc intestine is also reported. Two different bands, corresponding to the immature (27 KDa band) and the glycosilated isoform (30 KDa band), were detected. (Lower part) Confocal microscopy analysis of RSPO1 distribution observed by whole mount immunofluorescence (FITC signal) in 15.5 dpc ovary (positive control) and in 13.5 dpc male germ cells. RSPO1 immuno-labeled germ cells were double stained with anti-VASA (germ cell marker) antibody. Right panel: Confocal microscopy analysis of RSPO1 distribution observed by whole mount immunofluorescence (FITC signal) in 11.5 (A), 12.5 (C and D), 13.5 (E and F), 16.5 (G and H), and 18.5 dpc (I and L) male UGRs at different magnifications. In B the corresponding bright field of 11.5 dpc male genital ridge is reported. ce: coelomic epithelium; gr: genital ridge; ms: mesonephros; tc: testicular cords; ts: testis, ov: ovary. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0124213), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of R-Spondin 1 by Western Blot

Detection of R-Spondin 1 by Western Blot

Spatiotemporal expression of chicken RSPO1. (A) Relative mRNA expression of RSPO1 in tissues of E12 (HH 38) male and female chicken embryos. n = 4. (B) Relative mRNA expression pattern of RSPO1 at different stages of chicken embryos, E12 =12 days of incubation. Beta-actin was used as a reference gene. n = 4. (C) Relative protein expression of RSPO1 in male and female gonads of E12 chicken embryos. Data are shown as mean ± SEM. Tubulin was used as a reference gene. n = 5. (D) RSPO1 mRNA expression levels in left and right gonads of E12 chick embryos. (E) Localization of RSPO1 protein expression in the ovaries and testes of E12 chick embryos. Scale bars = 50 μm. M: medulla. C: cortex. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37444038), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of R-Spondin 1 by Western Blot

Detection of R-Spondin 1 by Western Blot

Effect of FAD treatment on the expression of RSPO1 in E12 chicken embryonic gonads. (A) Anatomy of male and female gonads of FAD-treated group and control group. L = left gonad, R = right gonad. Scale bars = 1cm. (B) Effect of FAD treatment on the mRNA expression of RSPO1 and some other genes in chicken embryonic gonads. Beta-actin was used as a reference gene. n = 4. (C) Effect of FAD treatment on the protein expression of RSPO1 in chicken embryonic gonads (the wrong symbol X indicates a false sample loading whose result was not included in the analysis). n = 4. Tubulin was used as a reference gene, and data are shown as mean ± SEM. FAD: fadrozole. FAD treatment was performed on E2.5 (embryonic day 2.5). E12 = embryonic day 12. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37444038), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of R-Spondin 1 by Immunohistochemistry

Detection of R-Spondin 1 by Immunohistochemistry

Spatiotemporal expression of chicken RSPO1. (A) Relative mRNA expression of RSPO1 in tissues of E12 (HH 38) male and female chicken embryos. n = 4. (B) Relative mRNA expression pattern of RSPO1 at different stages of chicken embryos, E12 =12 days of incubation. Beta-actin was used as a reference gene. n = 4. (C) Relative protein expression of RSPO1 in male and female gonads of E12 chicken embryos. Data are shown as mean ± SEM. Tubulin was used as a reference gene. n = 5. (D) RSPO1 mRNA expression levels in left and right gonads of E12 chick embryos. (E) Localization of RSPO1 protein expression in the ovaries and testes of E12 chick embryos. Scale bars = 50 μm. M: medulla. C: cortex. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37444038), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of R-Spondin 1 by Western Blot

Detection of R-Spondin 1 by Western Blot

Spatiotemporal expression of chicken RSPO1. (A) Relative mRNA expression of RSPO1 in tissues of E12 (HH 38) male and female chicken embryos. n = 4. (B) Relative mRNA expression pattern of RSPO1 at different stages of chicken embryos, E12 =12 days of incubation. Beta-actin was used as a reference gene. n = 4. (C) Relative protein expression of RSPO1 in male and female gonads of E12 chicken embryos. Data are shown as mean ± SEM. Tubulin was used as a reference gene. n = 5. (D) RSPO1 mRNA expression levels in left and right gonads of E12 chick embryos. (E) Localization of RSPO1 protein expression in the ovaries and testes of E12 chick embryos. Scale bars = 50 μm. M: medulla. C: cortex. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37444038), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of R-Spondin 1 by Western Blot

Detection of R-Spondin 1 by Western Blot

Effect of FAD treatment on the expression of RSPO1 in E12 chicken embryonic gonads. (A) Anatomy of male and female gonads of FAD-treated group and control group. L = left gonad, R = right gonad. Scale bars = 1cm. (B) Effect of FAD treatment on the mRNA expression of RSPO1 and some other genes in chicken embryonic gonads. Beta-actin was used as a reference gene. n = 4. (C) Effect of FAD treatment on the protein expression of RSPO1 in chicken embryonic gonads (the wrong symbol X indicates a false sample loading whose result was not included in the analysis). n = 4. Tubulin was used as a reference gene, and data are shown as mean ± SEM. FAD: fadrozole. FAD treatment was performed on E2.5 (embryonic day 2.5). E12 = embryonic day 12. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37444038), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of R-Spondin 1 by Immunohistochemistry

Detection of R-Spondin 1 by Immunohistochemistry

Spatiotemporal expression of chicken RSPO1. (A) Relative mRNA expression of RSPO1 in tissues of E12 (HH 38) male and female chicken embryos. n = 4. (B) Relative mRNA expression pattern of RSPO1 at different stages of chicken embryos, E12 =12 days of incubation. Beta-actin was used as a reference gene. n = 4. (C) Relative protein expression of RSPO1 in male and female gonads of E12 chicken embryos. Data are shown as mean ± SEM. Tubulin was used as a reference gene. n = 5. (D) RSPO1 mRNA expression levels in left and right gonads of E12 chick embryos. (E) Localization of RSPO1 protein expression in the ovaries and testes of E12 chick embryos. Scale bars = 50 μm. M: medulla. C: cortex. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37444038), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Mouse R-Spondin 1 Antibody

Application
Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed frozen sections of mouse ovary (E13.5)

Western Blot

0.1 µg/mL
Sample: Recombinant Mouse R-Spondin 1 (Catalog # 3474-RS)

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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.

Calculators

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 1

R-Spondin 1 (RSPO1, Roof plate-specific Spondin 1), also known as cysteine-rich and single thrombospondin domain containing protein 3 (Cristin 3), is a 27 kDa secreted protein that belongs to the R-Spondin family (1, 2). R-Spondins share around 40% aa identity. All regulate Wnt/ beta -catenin signaling, but have distinct expression patterns (1 - 3). Like other R-spondins, R-Spondin 1 contains two adjacent cysteine-rich furin-like domains (amino acids (aa) 34 - 135) followed by a thrombospondin (TSP-1) motif (aa 147 - 207) and a region rich in basic residues (aa 211 - 263). Only the furin-like domains are needed for beta -catenin stabilization (2, 4). A putative nuclear localization signal at the C-terminus may allow some expression in the nucleus (5). R-Spondin 1 contains one potential N-glycosylation site. Over aa 21 - 209, mouse R-Spondin 1 shares 98%, 94%, 94%, 93%, 92% and 88% aa identity with rat, human, horse, cow, goat and dog RSPO-1, respectively. R-Spondin 1 is expressed in early development at the roof plate boundary and is thought to contribute to dorsal neural tube development (3, 5). In humans, rare disruptions of the R-Spondin 1 gene are associated with tendencies for XX sex reversal (phenotypic male) or hermaphroditism, indicating a role for R-Spondin 1 in gender-specific differentiation (6, 7). Disruption is also associated with palmoplantar keratosis (6, 7). Postnatally, R-Spondin 1 is expressed by neuroendocrine cells in the intestine, adrenal gland and pancreas, and by epithelia in kidney and prostate (8). Injection of recombinant R-Spondin 1 in mice causes activation of beta -catenin and proliferation of intestinal crypt epithelial cells, and ameliorates experimental colitis (8, 9). R-Spondin 1 appears to regulate Wnt/ beta -catenin by competing with the Wnt antagonist DKK-1 for binding to the Wnt co-receptor, Kremen (10). This competition reduces internalization of DKK-1/LRP-6/Kremen complexes (10). Reports differ on whether R‑Spondin 1 binds LRP-6 directly (10 - 12).

References

  1. Lowther, W. et al. (2005) J. Virol. 79:10093.
  2. Kim, K-A. et al. (2006) Cell Cycle 5:23.
  3. Nam, J-S. et al. (2007) Gene Expr. Patterns 7:306.
  4. Kazanskaya, O. et al. (2004) Dev. Cell 7:525.
  5. Kamata, T. et al. (2004) Biochim. Biophys. Acta 1676:51.
  6. Tomaselli, S. et al. (2008) Hum. Mutat. 29:220.
  7. Parma, P. et al. (2006) Nat. Genet. 38:1304.
  8. Kim, K-A. et al. (2005) Science 309:1256.
  9. Zhao, J. et al. (2007) Gastroenterology 132:1331.
  10. Binnerts, M.E. et al. (2007) Proc. Natl. Acad. Sci. USA 104:14700.
  11. Nam, J-S. et al. (2006) J. Biol. Chem. 281:13247.
  12. Wei, Q. et al. (2007) J. Biol. Chem. 282:15903.

Long Name

Roof Plate-specific Spondin 1

Alternate Names

Cristin 3, CRISTIN3, HRspo1, RSPO, RSPO1, RSpondin 1

Entrez Gene IDs

284654 (Human); 192199 (Mouse); 102122369 (Cynomolgus Monkey)

Gene Symbol

RSPO1

UniProt

Q9Z132

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Product Documents for Mouse R-Spondin 1 Antibody

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Product Specific Notices for Mouse R-Spondin 1 Antibody

For research use only

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