Mouse Urocortin PharmaGenie ELISA Kit (SBRS0085)

1. Pre-Coated 96-well Strip Microplate
2. Wash Buffer
3. Standard Peptide
4. Assay Diluent(s)
5. Biotinylated Peptide
6. HRP-Streptavidin
7. TMB One-Step Substrate
8. Stop Solution
9. Assay Diagram
10. Positive Control Sample
11. Capture Antibody
12. Technical Manual

Other materials and equipment required:

The Assay Genie Mouse Urocortin PharmaGenie ELISA Kit (SBRS0085) will require other equipment and materials to carry out the assay. Please see list below for further details.

• Distilled or deionized water
• Precision pipettes to deliver 2 ul to 1 ml volumes
• Adjustable 1-25 ml pipettes for reagent preparation
• 100 ml and 1 liter graduated cylinders
• Tubes to prepare standard and sample dilutions
• Orbital shaker
• Aluminum foil
• Saran Wrap
• Absorbent paper
• Microplate reader capable of measuring absorbance at 450nm
• SigmaPlot software (or other software that can perform four-parameter logistic regression models)

1. Prepare all reagents, samples and standards as instructed.
2. Add 100 ul detection antibody to each well.
3. Incubate 1.5 h at RT or O/N at 4°C.
4. Add 100 ul standard or sample to each well.
5. Incubate 2.5 h at RT.
6. Add 100 ul prepared streptavidin solution.
7. Incubate 45 min at RT.
8. Add 100 ul TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 ul Stop Solution to each well.
11. Read plate at 450 nm immediately.

Urocortin is a member of the sauvagine/corticotropin-releasing factor/urotensin I family. It is an endogenous ligand for corticotropin-releasing factor (CRF) receptors. Urocortin, peptide of 40 amino acids, acts in vitro to stimulate the secretion of adrenocorticotropic hormone (ACTH). It binds with high affinity to CRF receptor types 1, 2-alpha, and 2-beta. Urocortin plays a role in the establishment of normal hearing thresholds. It reduces food intake and regulates ghrelin levels in gastric body and plasma. Urocortin is a potent anorexigenic peptide that induces fed-like motor activity when administered centrally or peripherally in fasted animals. Urocortin is also a potent and long-lasting hypotensive agent and increases coronary blood flow.