Human Substance P PharmaGenie ELISA Kit (SBRS0173)

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 Human Substance P PharmaGenie ELISA Kit (SBRS0173) 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.

Substance P (SP) is an 11 amino acid peptide produced from a precursor protein, Protachykinin-1. It is a neuropeptide, acting as a neurotransmitter and as a neuromodulator. It is found in the brain and spinal cord and is associated with inflammatory processes and pain. Its receptor, NK1R, is distributed over cytoplasmic and nuclear membranes of many cell types (neurons, glia, endothelia of capillaries and lymphatics, fibroblasts, stem cells, white blood cells) in many tissues and organs. Substance P has been found as a key first responder to most noxious/extreme stimuli (stressors). It has been reported that Substance P is involved in many biological events, such as vasodilation, inflammation, pain, mood, anxiety, Learning, vomiting, cell growth, proliferation, angiogenesis, and migration. Elevation of Substance P and/or its receptor (NK1R) in serum, plasma, or tissue has been associated with many diseases.