Substance P Parameter Assay Kit

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Multi-species Substance P ELISA Standard Curve
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Product Details
Citations (34)
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Substance P Parameter Assay Kit Summary

96-well strip plate
Assay Length
3.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (50 uL), Heparin Plasma (50 uL), Saliva (50 uL), Urine (50 uL)
43.8 pg/mL
Assay Range
39.0 - 2,500 pg/mL (Cell Culture Supernates)
Measures Substance P levels in various samples
Cross-reactivity observed with 1 or more available related molecules.Cross-species reactivity not tested.
No significant interference observed with available related molecules.

Product Summary

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.


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


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.
Multi-species Substance P ELISA Linearity

Scientific Data

Multi-species Substance P ELISA Standard Curve

Product Datasheets

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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.

Entrez Gene IDs:
6863 (Human)
Alternate Names:
neurokinin 1; neurokinin 2; neurokinin A; Neurokinin-1; neuromedin L, neurokinin alpha, neuropeptide K, neuropeptide gamma); neuropeptide gamma; neuropeptide K; NK2; NKA; NKNAneurokinin alpha; NPK; PPT; protachykinin-1; substance K; Substance P; TAC1; TAC2neuromedin L; tachykinin 2; tachykinin, precursor 1 (substance K, substance P, neurokinin 1, neurokinin 2; tachykinin, precursor 1
⚠ 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

Assay Procedure

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.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. Calibrator Diluent
  4.   Add 100 µL of Calibrator Diluent to the non-specific binding (NSB) wells.
  5.   Add 50 µL of Calibrator Diluent to the zero standard (B0) wells.

  6. 50 µL Standard, Control, or Sample
  7.   Add 50 µL of Standard, Control, or sample to the remaining wells.

  8. 50 µL Primary Antibody Solution
  9.   Add 50 µL of Primary Antibody Solution to each well (except the NSB wells).

  10. 50 µL Conjugate
  11.   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.
  12.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  13. 200 µL Substrate Solution
  14.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  15. 50 µL Stop Solution
  16. 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.

Citations for Substance P Parameter Assay Kit

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.

34 Citations: Showing 1 - 10
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  1. Interleukin-10 signaling in somatosensory neurons controls CCL2 release and inflammatory response
    Authors: de Souza, S;Rosario Claudio, J;Sim, J;Inyang, KE;Dagenais, A;Monahan, K;Lee, B;Ramakrishnan, H;Parmar, V;Geron, M;Scherrer, G;Folger, JK;Laumet, G;
    Brain, behavior, and immunity
    Species: Mouse
    Sample Types: Cell Culture Supernates
  2. Tracheobronchial-esophageal reflex initiates esophageal hypersensitivity and aggravates cough hyperreactivity in guinea pigs with esophageal acid infusion
    Authors: Q Chen, R Dong, L Zhang, M Zhang, X Xu, L Yu, Z Qiu
    Respiratory physiology & neurobiology, 2022-03-28;301(0):103890.
    Species: Guinea Pig
    Sample Types: BALF
  3. Analgesic effects of a 5-HT3 receptor antagonist in an animal model of complex regional pain syndrome
    Authors: JH Park, CH Lee, HD Ham, ES Choi, C Lee, S Lee
    European review for medical and pharmacological sciences, 2021-11-01;25(22):7051-7057.
    Species: Rats
    Sample Types: Plasma
  4. Cough hypersensitivity in patients with metabolic syndrome: a clinical finding and its possible mechanisms
    Authors: J Cheng, Z Xie, S Wang, S Wen, S Niu, C Shi, L Yu, X Xu
    BMC pulmonary medicine, 2021-09-06;21(1):284.
    Species: Human
    Sample Types: Sputum Supernates
  5. NK1 antagonists attenuate tau phosphorylation after blast and repeated concussive injury
    Authors: F Corrigan, I Cernak, K McAteer, SC Hellewell, JV Rosenfeld, RJ Turner, R Vink
    Scientific Reports, 2021-04-23;11(1):8861.
    Species: Rat
    Sample Types: Tissue Homogenates
  6. Intravenous allogeneic umbilical cord blood-derived mesenchymal stem cell therapy in recessive dystrophic epidermolysis bullosa patients
    Authors: SE Lee, SJ Lee, SE Kim, K Kim, B Cho, K Roh, SC Kim
    JCI Insight, 2021-01-25;6(2):.
    Species: Human
    Sample Types: Serum
  7. Elevated lead levels in relation to low serum neuropeptide Y and adverse behavioral effects in preschool children with e-waste exposure
    Authors: X Zeng, C Xu, X Xu, Y Zhang, Y Huang, X Huo
    Chemosphere, 2020-12-21;269(0):129380.
    Species: Human
    Sample Types: Serum
  8. Comparison of tear proteomic and neuromediator profiles changes between small incision lenticule extraction (SMILE) and femtosecond laser-assisted in-situ keratomileusis (LASIK)
    Authors: YC Liu, GH Yam, MT Lin, E Teo, SK Koh, L Deng, L Zhou, L Tong, JS Mehta
    Journal of advanced research, 2020-11-05;29(0):67-81.
    Species: Human
    Sample Types: Tear Supernates
  9. Effects of a Forest Walk on Urinary Dityrosine and Hexanoyl-Lysine in Young People: A Pilot Study
    Authors: A Yamada, Y Sato, T Horike, M Miyanaga, DH Wang
    Int J Environ Res Public Health, 2020-07-10;17(14):.
    Species: Human
    Sample Types: Urine
  10. Bilateral Effect of the Unilateral Corneal Nerve Cut on Both Ocular Surface and Lacrimal Gland
    Authors: HK Lee, KW Kim, JS Ryu, HJ Jeong, SM Lee, MK Kim
    Invest. Ophthalmol. Vis. Sci., 2019-01-02;60(1):430-441.
    Species: Mouse
    Sample Types: Tissue Homogenates
  11. Tachykinin NK1 receptor antagonist L-733,060 and substance P deletion exert neuroprotection through inhibiting oxidative stress and cell death after traumatic brain injury in mice
    Authors: Q Li, X Wu, Y Yang, Y Zhang, F He, X Xu, Z Zhang, L Tao, C Luo
    Int. J. Biochem. Cell Biol., 2018-12-26;0(0):.
    Species: Mouse
    Sample Types: Serum
  12. TRPA1 and substance P mediate stress induced duodenal lesions in water immersion restraint stress rat model
    Authors: Y Xu, C Huang, H Deng, J Jia, Y Wu, J Yang, W Tu
    Turk J Gastroenterol, 2018-11-01;29(6):692-700.
    Species: Mouse
    Sample Types: Tissue Homogenates
  13. Neurokinin-1 receptor antagonism improves postoperative neurocognitive disorder in mice.
    Authors: Li Z, Luo T, Ning X, Xiong C, Wu A
    Neurosci Lett, 2018-09-28;687(0):189-195.
    Species: Mouse
    Sample Types: Tissue Homogenates
  14. Neuroepithelial control of mucosal inflammation in acute cystitis
    Authors: DSC Butler, I Ambite, K Nagy, C Cafaro, A Ahmed, A Nadeem, N Filenko, TH Tran, KE Andersson, B Wullt, M Puthia, C Svanborg
    Sci Rep, 2018-07-20;8(1):11015.
    Species: Mouse
    Sample Types: Urine
  15. Sustained Low Serum Substance P Levels in Non-Surviving Septic Patients
    Authors: L Lorente, MM Martín, A Pérez-Ceja, J Ferreres, J Solé-Violá, L Labarta, C Díaz, A Jiménez
    Int J Mol Sci, 2017-07-15;18(7):.
    Species: Human
    Sample Types: Serum
  16. Curcumin alleviates lumbar radiculopathy by reducing neuroinflammation, oxidative stress and nociceptive factors
    Authors: L Xiao, M Ding, A Fernandez, P Zhao, L Jin, X Li
    Eur Cell Mater, 2017-05-09;33(0):279-293.
    Species: Mouse
    Sample Types: Tissue Homogenates
  17. Genetic and pharmacological antagonism of NK1 receptor prevents opiate abuse potential
    Authors: AJ Sandweiss, MI McIntosh, A Moutal, R Davidson-K, J Hu, AK Giri, T Yamamoto, VJ Hruby, R Khanna, TM Largent-Mi, TW Vanderah
    Mol. Psychiatry, 2017-05-09;0(0):.
    Species: Rat
    Sample Types: CSF
  18. The dual regulation of substance P-mediated inflammation via human synovial mast cells in rheumatoid arthritis
    Authors: Y Okamura, S Mishima, JI Kashiwakur, T Sasaki-Sak, S Toyoshima, K Kuroda, S Saito, Y Tokuhashi, Y Okayama
    Allergol Int, 2017-03-31;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  19. Cytokine and neuropeptide levels are associated with pain relief in patients with chronically painful total knee arthroplasty: a pilot study
    Authors: JA Singh, S Noorbalooc, KL Knutson
    BMC Musculoskelet Disord, 2017-01-14;18(1):17.
    Species: Human
    Sample Types: Serum
  20. Sphenopalatine Ganglion Acupuncture Improves Nasal Ventilation and Modulates Autonomic Nervous Activity in Healthy Volunteers: A Randomized Controlled Study
    Sci Rep, 2016-07-18;6(0):29947.
    Species: Human
    Sample Types: Tissue Secretion
  21. Efficacy of oral meloxicam suspension for prevention of pain and inflammation following band and surgical castration in calves
    Authors: M E Olson
    BMC Vet Res, 2016-06-13;12(1):102.
    Species: Bovine
    Sample Types: Plasma
  22. Brain injury induces specific changes in the caecal microbiota of mice via altered autonomic activity and mucoprotein production
    Authors: A Denes
    Brain Behav. Immun., 2016-04-06;57(0):10-20.
    Species: Mouse
    Sample Types: Tissue Homogenates
  23. TRPA1 controls inflammation and pruritogen responses in allergic contact dermatitis.
    Authors: Liu, Boyi, Escalera, Jasmine, Balakrishna, Shrilath, Fan, Lu, Caceres, Ana I, Robinson, Eve, Sui, Aiwei, McKay, M Craig, McAlexander, M Allen, Herrick, Christin, Jordt, Sven E
    FASEB J, 2013-05-30;27(9):3549-63.
    Species: Mouse
    Sample Types: Tissue Homogenates
  24. The novel cytokine interleukin-33 activates acinar cell proinflammatory pathways and induces acute pancreatic inflammation in mice.
    Authors: Kempuraj D, Twait E, Williard D, Yuan Z, Meyerholz D, Samuel I
    PLoS ONE, 2013-02-13;8(2):e56866.
    Species: Mouse, Rat
    Sample Types: Tissue Homogenates
  25. Prophylactic and Therapeutic Targeting of the Neurokinin-1 Receptor Limits Neuroinflammation in a Murine Model of Pneumococcal Meningitis.
    Authors: Chauhan VS, Kluttz JM
    J. Immunol., 2011-05-11;0(0):.
    Species: Mouse
    Sample Types: Tissue Homogenates
  26. The change of cytokines in tear and blood after different pterygium operation.
    Authors: Lee JK, Song YS, Shin JS, Kwon YS, Shin MS, Kim JC
    Cytokine, 2009-12-08;49(2):148-54.
    Species: Human
    Sample Types: Tears
  27. Role of sensory innervation in the rat pulmonary neutrophil recruitment induced by staphylococcal enterotoxins type A and B.
    Authors: Desouza IA, Camargo EA, Mariano NS, Optiz-Neto JB, Resende JS, Mello GC, Costa SK, De Nucci G, Antunes E
    Eur. J. Pharmacol., 2009-04-16;613(1):128-34.
    Species: Rat
    Sample Types: BALF
  28. A constitutively active form of neurokinin 1 receptor and neurokinin 1 receptor-mediated apoptosis in glioblastomas.
    Authors: Akazawa T, Kwatra SG, Goldsmith LE, Richardson MD, Cox EA, Sampson JH, Kwatra MM
    J. Neurochem., 2009-03-11;109(4):1079-86.
    Species: Human
    Sample Types: Cell Culture Supernates
  29. Chemical sympathectomy increases susceptibility to ocular herpes simplex virus type 1 infection.
    Authors: Templeton A, Nguyen G, Ash JD, Straub RH, Carr DJ
    J. Neuroimmunol., 2008-05-20;197(1):37-46.
    Species: Mouse
    Sample Types: Tissue Homogenates
  30. Basic fibroblast growth factor accelerates matrix degradation via a neuro-endocrine pathway in human adult articular chondrocytes.
    Authors: Im HJ, Li X, Muddasani P, Kim GH, Davis F, Rangan J, Forsyth CB, Ellman M, Thonar EJ
    J. Cell. Physiol., 2008-05-01;215(2):452-63.
    Species: Human
    Sample Types: Synovial Fluid
  31. Spinal CCL2 pronociceptive action is no longer effective in CCR2 receptor antagonist-treated rats.
    Authors: Dansereau MA, Gosselin RD, Pohl M, Pommier B, Mechighel P, Mauborgne A, Rostene W, Kitabgi P, Beaudet N, Sarret P, Melik-Parsadaniantz S
    J. Neurochem., 2008-04-17;106(2):757-69.
    Species: Rat
    Sample Types: Cell Lysates
  32. Association of cathepsin E deficiency with the increased territorial aggressive response of mice.
    Authors: Shigematsu N, Fukuda T, Yamamoto T, Nishioku T, Yamaguchi T, Himeno M, Nakayama KI, Tsukuba T, Kadowaki T, Okamoto K, Higuchi S, Yamamoto K
    J. Neurochem., 2008-01-21;105(4):1394-404.
    Species: Mouse
    Sample Types: Tissue Homogenates
  33. Contributions of histamine, prostanoids, and neurokinins to edema elicited by edema toxin from Bacillus anthracis.
    Authors: Tessier J, Green C, Padgett D, Zhao W, Schwartz L, Hughes M, Hewlett E
    Infect. Immun., 2007-01-29;75(4):1895-903.
    Species: Human
    Sample Types: Cell Culture Supernates
  34. Activity-dependent release of precursor nerve growth factor, conversion to mature nerve growth factor, and its degradation by a protease cascade.
    Authors: Bruno MA, Cuello AC
    Proc. Natl. Acad. Sci. U.S.A., 2006-04-17;103(17):6735-40.
    Species: Rat
    Sample Types: Whole Tissue


  1. Why is aprotinin used in this kit and is it necessary?

    • 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.

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Substance P Parameter Assay Kit
By Zhenping Wang on 09/30/2020
Sample Tested: cell culture supernatant from human epidermal keratinocytes