Human Pro‑Relaxin‑1 Antibody

Catalog #: AF3257 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF3257
AF3257-SP

Detection of Human Relaxin‑1 by Western Blot.

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All Relaxin-1 Antibodies

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human Pro‑Relaxin‑1 Antibody Summary

Species Reactivity

Human

Specificity

Detects human Relaxin-1 in direct ELISAs and Western blots. In direct ELISAs, less than 5% cross-reactivity with recombinant human (rh) Relaxin‑2 and rhRelaxin-3 is observed.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant human Relaxin-1
Lys26-Cys185
Accession # P04808

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.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

See below

ELISA

This antibody functions as an ELISA detection antibody when paired with Rat Anti-Human Relaxin‑1 Monoclonal Antibody (Catalog # MAB32571).

This product is intended for assay development on various assay platforms requiring antibody pairs. We recommend the Human Relaxin-1 DuoSet ELISA Kit (Catalog # DY3257) for convenient development of a sandwich ELISA.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Human Relaxin‑1 by Western Blot. Western blot shows lysates of LNCaP human prostate cancer cell line. PVDF Membrane was probed with 1 µg/mL of Goat Anti-Human Pro-Relaxin-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3257) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for Relaxin-1 at approximately 7 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

ELISA

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Human Relaxin‑1 ELISA Standard Curve. Recombinant Human Relaxin‑1 protein was serially diluted 2-fold and captured by Rat Anti-Human Relaxin‑1 Monoclonal Antibody (MAB32571) coated on a Clear Polystyrene Microplate (DY990). Goat Anti-Human Pro Relaxin‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3257) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (DY998) followed by Substrate Solution (DY999) and stopping the enzymatic reaction with Stop Solution (DY994).

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile 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: Relaxin-1

Human Relaxin-1, also called H1 Relaxin or RLN1, is one of three human relaxins in the structurally related insulin/relaxin superfamily (1, 2). Relaxin-1 is thought to be the result of duplication of the Relaxin-2 gene in higher primates only. In species below higher primates, Relaxin-1 is the equivalent of human Relaxin-2. Relaxin-1 is found in some but not all tissues expressing Relaxin-2. It is prominent in the prostate, but also present in decidua, placenta, endometrium and at low levels in the myocardium (2, 3). As with other insulin/relaxin superfamily members, human Relaxin-1 is synthesized as a preprohormone (4). Processing of the 21 kDa preprorelaxin-1 includes removal of the signal sequence, formation of two disulfide bonds between A and B chains and removal of the intervening C-chain by a prohormone convertase. The resulting mature protein is an unglycosylated, 6 kDa dimer of disulfide-linked A and B chains. Human Relaxin-1 shares 76% amino acid (aa) identity with human Relaxin-2, and 43%, 50%, and 43% aa identity with mouse, rat, and canine Relaxin-1, respectively. An alternate splice form of unknown significance has a 47 aa substitution which does not have typical C-chain cleavage motifs (5). Relaxins confer activity by binding to leucine-rich G-protein coupled receptors LGR7 and LGR8 (2, 6). Prostatic relaxins are anti-apoptotic and contribute to development and maintenance of male fertility. It is not clear whether human Relaxins -1 and -2 have distinct functions. Both use the same receptor and have the same critical amino acids for folding and for receptor interaction. While receptor affinity is similar, activity is lower for Relaxin-1 as compared to Relaxin-2 (7). Progesterone increases expression of only Relaxin-2, while glucocorticoids increase expression of both (8).

References