Human Relaxin-3 (H3 relaxin, INSL7) is one of seven relaxin-like peptides belonging to the insulin superfamily (1‑4). Unlike human relaxins 1 and 2, it does not play a role in reproduction but appears to be a neuropeptide involved in stress response in the brain stem (3‑5). The single-chain human Prorelaxin-3 shares 83% and 80% amino acid (aa) sequence identity with mouse and rat prorelaxin-3, respectively.The 142 aa Relaxin-3 pre-proprotein is processed to remove a 25 aa signal peptide and a connecting peptide (aa 53‑118). The resulting mature Relaxin-3 is a 5.5 kDa, 51 aa secreted heterodimer of A (aa 119‑142) and B (aa 26‑52) peptides connected by two intermolecular disulfide bonds (1). Mature human Relaxin-3 is 96%, 94%, and 92% aa identical to porcine, canine, and mouse Relaxin-3, respectively. This is much higher identity between species than that seen for other relaxins. Relaxin-3 is thus suggested to be the ancestral relaxin family member (2). Relaxin-3 is the only known ligand for the G-protein-coupled receptor GPCR135, designated RXFP3 (4, 6). In rodents, GPCR135 is expressed primarily in the supraoptic and paraventricular nucleus (6). This region has connections to the dorsal tegmental region of the pons (also called the nucleus incertus), where expression of Relaxin-3 is highest (5). Relaxin-3 also binds the more widely-expressed LGR7 (RXFP1) receptor, but with lower affinity than that of Relaxin-2 (1, 7). Although binding of Relaxin-3 to LGR7 increases intracellular cAMP, binding to GPCR135 inhibits cAMP accumulation, indicating coupling to Gi, Go or Gz by this receptor (1, 5). Relaxin-3 expression does not overlap well with its other receptor, GPCR142, which instead appears to be the primary receptor for INSL5 (3, 8).
Key Product Details
Species Reactivity
Human
Applications
Western Blot, Neutralization
Label
Unconjugated
Antibody Source
Monoclonal Mouse IgG2B Clone # 332105
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Product Specifications
Immunogen
E. coli-derived recombinant human Relaxin-3
Arg26-Cys142
Accession # Q8WXF3
Arg26-Cys142
Accession # Q8WXF3
Specificity
Recognizes the pro and mature forms of human Relaxin-3 in direct ELISAs and Western blots. In direct ELISAs and Western blots, no cross‑reactivity with the pro forms of recombinant human (rh) Relaxin-1 or rhRelaxin-2 is observed.
Clonality
Monoclonal
Host
Mouse
Isotype
IgG2B
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.
Scientific Data Images for Human Relaxin‑3 Antibody
cAMP production Induced by Relaxin‑3 and Neutralization by Human Relaxin‑3 Antibody.
Recombinant Human Relaxin-3 (Catalog # 3107-RN) induces cAMP production in the THP-1 human acute monocytic leukemia cell line in a dose-dependent manner (orange line), as measured by cAMP Parameter Assay Kit (Catalog # KGE002B). cAMP production elicited by Recombinant Human Relaxin-3 (25 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human Relaxin-3 Monoclonal Antibody (Catalog # MAB31071). The ND50 is typically 1-5 µg/mL.Applications for Human Relaxin‑3 Antibody
Application
Recommended Usage
Western Blot
1 µg/mL
Sample: Recombinant Human Relaxin‑3 (Catalog # 3107-RN) under non-reducing conditions only
Sample: Recombinant Human Relaxin‑3 (Catalog # 3107-RN) under non-reducing conditions only
Neutralization
Measured by its ability to neutralize Relaxin‑3-induced cAMP production in the THP‑1 human acute monocytic leukemia cell line. Parsell, D. A. et al. (1996) J. Biol. Chem. 271:27936. The Neutralization Dose (ND50) is typically 1-5 µg/mL in the presence of 25 ng/mL Recombinant Human Relaxin‑3.
Formulation, Preparation, and Storage
Purification
Protein A or G purified from hybridoma culture supernatant
Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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Formulation
Lyophilized from a 0.2 μm filtered solution in PBS and NaCl 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
Background: Relaxin-3
References
- Kizawa, H. et al. (2003) Regul. Pept. 113:79.
- Wilkinson, T.N. et al. (2005) BMC Evol. Biol. 5:14.
- Bathgate, R.A. et al. (2006) Pharmacol. Rev. 58:7.
- Liu, C. et al. (2005) Ann. N.Y. Acad. Sci. 1041:47.
- Tanaka, M. et al. (2005) Eur. J. Neurosci. 21:1659.
- Liu, C. et al. (2003) J. Biol. Chem. 278:50754.
- Rosengren, K.J. et al. (2006) J. Biol. Chem. 281:5845.
- Liu, C. et al. (2003) J. Biol. Chem. 278:50765.
Alternate Names
H3, INSL7, M3, Relaxin3, RLN3, RXN3, ZINS4
Entrez Gene IDs
117579 (Human)
Gene Symbol
RLN3
UniProt
Additional Relaxin-3 Products
Product Documents for Human Relaxin‑3 Antibody
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human Relaxin‑3 Antibody
For research use only
Related Research Areas
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Cellular Response to Hypoxia Protocols
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- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
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- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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