Mouse Secretin PharmaGenie ELISA Kit (SBRS0080)

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 Secretin PharmaGenie ELISA Kit (SBRS0080) 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.

Secretin is a 27 amino acids peptide that is cleaved from a 120 amino acid precursor protein encoded by the SCT gene in human. Secretin is synthesized in cytoplasmic secretory granules of S-cells, which are found mainly in the mucosa of the duodenum, and in smaller numbers in the jejunum of the small intestine. Secretin is a peptide hormone that regulates water homeostasis throughout the body, and influences the environment of the duodenum by regulating secretions in the stomach and pancreas. Secretin also helps regulate the pH of the duodenum by: inhibiting the secretion of gastric acid from the parietal cells of the stomach; and stimulating the production of bicarbonate from the centroacinar cells and intercalated ducts of the pancreas. In 2007, secretin was discovered to play a role in osmoregulation by acting on the hypothalamus, pituitary, and kidney. Secretin has been widely used in medical field especially in pancreatic functioning test because it increases pancreatic secretions. This test can provide information about whether there are any abnormalities in the pancreas which can include gastrinoma, pancreatitis or pancreatic cancer. Secretin has been proposed as a possible treatment for autism based on a hypothetical gut-brain connection; as yet there is no evidence to support it as effective.