Mouse Oxytocin EasyTest™ EIA Kit

Product Features

Product Name            Mouse Oxytocin EasyTest™ EIA Kit
Assay Type                   Solid Phase Competitive ELISA
Format                           96-well strip plate
Assay Process             Two wash step, three incubations
Assay Length              2.5 hours
Sample Volume         Serum , Plasma, 50µL
Detection Method    Colorimetric
Sensitivity                   80pg/mL
Assay Range                0.024-100ng/mL
Reactivity                     Mouse
Cross-Reactivity       No cross-reactivity observed with available related molecules
Gene Names               Oxt; OT; Oxy
Gene ID                         18429
Swissprot ID               P35454

Standard Curve        Mouse Oxytocin EasyTest™ EIA Kit

Kit Components

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Stop Solution
• Assay Diluent
• Anti-Oxytocin Antibody
• Standard Oxytocin peptide
• Biotinylated Oxytocin peptide
• TMB One-Step Substrate

Protocol Outline

1. Prepare all reagents, samples and standards as instructed in the manual.
2. Add 25 µl of anti-Oxytocin antibody to all wells.
3. Add 75 µl of 1 x assay diluent to the blank wells and 50 µl to B0 wells.
4. Add 50 µl of standard peptide/samples to the appropriate wells.
5. Add 25 µl of biotinylated peptide into each well except the blank wells.
6. Incubate 1.5 h at RT.
7. Wash and add 100 µl of diluted streptavidin-HRP solution to all wells.
8. Incubate 45 min at RT.
9. Wash and add 100 µl of TMB Substrate Solution into each well.
10. Incubate 15 min at RT.
11. Add 50 µl of Stop Solution to each well.
12. Read at 450 nm/570 nm.

Application

In blood, oxytocin binds with big proteins. Acidification of blood is required to separate and purify oxytocin.

Background

Oxytocin (OXT) is a mammalian neurohypophysial hormone. Produced by the hypothalamus and stored and secreted by the posterior pituitary gland. Oxytocin is a peptide of nine amino acids  with the sequence of cys –tyr – Ile – gln – asn – cys – pro – leu – gly. The oxytocin peptide is synthesized as an inactive precursor protein from the OXT gene. Its actions are mediated by a G-protein coupled receptor that requires Mg2+ and cholesterol. It belongs to the rhodopsin-type group of G-protein-coupled receptors. Oxytocin plays an important role in the neuroanatomy of intimacy, specifically in sexual reproduction of both sexes, in particular during and after childbirth; It is released in large amounts after distension of the cervix and uterus during labor, facilitating birth, maternal bonding, and, after stimulation of the nipples, lactation.

Storage

The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.

Regulatory Status      

Mouse Oxytocin EasyTest™ EIA Kit is for Research Use Only.

• Manual
• AssayProcedure
• MSDS