Cross-reactivity observed with 1 or more available related molecules.Cross-species reactivity not tested.
No significant interference observed with available related molecules.
The Parameter Substance P Immunoassay is a 3.5 hour competitive enzyme immunoassay designed to measure Substance P in cell culture supernates, serum, plasma, saliva, and urine. It contains a synthetically derived human Substance P peptide and has been shown to accurately quantitate this peptide. Results obtained using samples containing natural Substance P showed linear curves that were parallel to the standard curves obtained using the Parameter kit standards. These results indicate that this kit can be used to determine relative mass values for natural Substance P.
Intra-Assay Precision (Precision within an assay) Four samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Four samples of known concentration were tested in separate assays to assess inter-assay precision.
The recovery of Substance P spiked to levels throughout the range of the assay in various matrices was evaluated.
Average % Recovery
Cell Culture Media (n=4)
Heparin Plasma (n=4)
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of Substance P were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.
Background: Substance P
Substance P (SP, Neurokinin-1), is member of the Tachykinin peptide family and binds primarily to the Neurokinin 1 Receptor (NK1R, Substance P receptor, Tachykinin receptor 1) and with low affinity to the NK2 and NK3 receptors. Substance P is processed from preproTachykinin a, b, g and d isoforms that also generate other Tachykinins Neuropeptide K, Neurokinin A, Neurokinin B, and Neuropeptide G. Substance P is produced in the central nervous system, primary sensory neurons (C-type fibers), and endocrine cells of the intestinal mucosa. It triggers pain signals in the spinal cord dorsal horn as well as hypotension, tachycardia, release of inflammatory mediators, and direct antimicrobial action.
Refer to the product for complete assay procedure.
Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
Prepare all reagents, standard dilutions, and samples as directed in the product insert.
Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
Add 100 µL of Calibrator Diluent to the non-specific binding (NSB) wells.
Add 50 µL of Calibrator Diluent to the zero standard (B0) wells.
50 µL Standard, Control, or Sample
Add 50 µL of Standard, Control, or sample to the remaining wells.
50 µL Primary Antibody Solution
Add 50 µL of Primary Antibody Solution to each well (except the NSB wells).
50 µL Conjugate
Add 50 µL of Conjugate to each well. Cover with a plate sealer, and incubate at room temperature for 3 hours on a horizontal orbital microplate shaker.
Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
200 µL Substrate Solution
Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
50 µL Stop Solution
Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
R&D Systems personnel manually curate a database that contains references using R&D Systems products.
The data collected includes not only links to publications in PubMed,
but also provides information about sample types, species, and experimental conditions.