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.
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Substance P Parameter Assay Kit
R&D Systems | Catalog # KGE007
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Key Product Details
Assay Length
3.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Serum (50 µL), Heparin Plasma (50 µL), Saliva (50 µL), Urine (50 µL)
Sensitivity
43.8 pg/mL
Assay Range
78.1-2500 pg/mL (Cell Culture Supernates)
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Product Summary for Substance P Parameter Assay Kit
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.
Product Specifications
Format
96-well strip plate
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
Biotin
Species
Multi-Species
Specificity
Measures Substance P levels in various samples
Cross-reactivity
Cross-reactivity observed with 1 or more available related molecules. Cross-species reactivity not tested.
Interference
No significant interference observed with available related molecules.
Precision
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.
Cell Culture Supernates, Heparin Plasma, Saliva, Serum, Urine
| Intra-Assay Precision | Inter-Assay Precision | |||||||
|---|---|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 4 | 1 | 2 | 3 | 4 |
| n | 20 | 20 | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 146 | 258 | 616 | 1973 | 140 | 262 | 684 | 1923 |
| Standard Deviation | 12.3 | 15.1 | 21.5 | 142 | 18.1 | 26.8 | 103 | 180 |
| CV% | 8.4 | 5.8 | 3.5 | 7.2 | 12.9 | 10.2 | 15.0 | 9.3 |
Recovery for Substance P Parameter Assay Kit
The recovery of Substance P spiked to levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=4) | 94 | 82-110 |
| Heparin Plasma (n=4) | 100 | 85-116 |
| Saliva (n=4) | 102 | 90-110 |
| Serum (n=4) | 101 | 85-117 |
| Urine (n=4) | 89 | 76-107 |
Linearity
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.
Scientific Data Images for Substance P Parameter Assay Kit
Multi-species Substance P ELISA Standard Curve
Preparation and Storage
Shipping
The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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
Additional Substance P Products
Product Documents for Substance P Parameter Assay Kit
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Substance P Parameter Assay Kit
For research use only
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.Related Research Areas
Citations for Substance P Parameter Assay Kit
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Sample Tested: cell culture supernatant from human epidermal keratinocytesVerified Customer | Posted 09/30/2020
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Protocols
View specific protocols for Substance P Parameter Assay Kit (KGE007):
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.
- Add 50 µL of Standard, Control, or sample to the remaining wells.
- Add 50 µL of Primary Antibody Solution to each well (except the NSB wells).
- 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.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
- 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.
Calibrator Diluent
50 µL Standard, Control, or Sample
50 µL Primary Antibody Solution
50 µL Conjugate
200 µL Substrate Solution
50 µL Stop Solution
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
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FAQs for Substance P Parameter Assay Kit
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Q: Why is aprotinin used in this kit and is it necessary?
A: Aprotinin is a protease inhibitor used in the preparation of serum and plasma samples for testing in this kit. Serum and plasma sample performance without aprotinin has not been evaluated in this kit.