Substance P Parameter Assay Kit

(19 citations)   
  • Format
    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)
  • Sensitivity
    43.8 pg/mL
  • Assay Range
    39.0 - 2,500 pg/mL (Cell Culture Supernates)
  • 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.
Control Available
QC163, Parameter Immunoassay ControlSet 657 for Substance P - Please Inquire
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.

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, Serum, Heparin Plasma, Saliva, Urine
Intra-Assay Precision Inter-Assay Precision
Sample12341234
n2020202020202020
Mean14625861619731402626841923
Standard Deviation12.315.121.514218.126.8103180
CV%8.45.83.57.212.910.2159.3

Recovery

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.
Substance P Parameter Assay Kit
Preparation and Storage
  • 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.
    • 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; TAC2neuromedin L; tachykinin 2; tachykinin, precursor 1 (substance K, substance P, neurokinin 1, neurokinin 2; tachykinin, precursor 1;
    Related Research Areas
    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:

    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.

    19 Citations: Showing 1 - 10
    Filter your results:

    Species
    Sample Type
    1. 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;18(7):.
      Species: Human
      Sample Type: Serum
    2. 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;33(0):279-293.
      Species: Mouse
      Sample Type: Tissue Homogenates
    3. 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;0(0):.
      Species: Rat
      Sample Type: CSF
    4. 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;0(0):.
      Species: Human
      Sample Type: Cell Culture Supernates
    5. 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;18(1):17.
      Species: Human
      Sample Type: Serum
    6. Sphenopalatine Ganglion Acupuncture Improves Nasal Ventilation and Modulates Autonomic Nervous Activity in Healthy Volunteers: A Randomized Controlled Study
      Sci Rep, 2016;6(0):29947.
      Species: Human
      Sample Type: Tissue Secretion
    7. 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;12(1):102.
      Species: Bovine
      Sample Type: Plasma
    8. 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;8(2):e56866.
      Species: Mouse
      Sample Type: Tissue Homogenates
    9. 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;27(9):3549-63.
      Species: Mouse
      Sample Type: Tissue Homogenates
    10. 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;0(0):.
      Species: Mouse
      Sample Type: Tissue Homogenates
    11. 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, 2010;49(2):148-54.
      Species: Human
      Sample Type: Tears
    12. 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;109(4):1079-86.
      Species: Human
      Sample Type: Cell Culture Supernates
    13. 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;613(1):128-34.
      Species: Rat
      Sample Type: BALF
    14. 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;197(1):37-46.
      Species: Mouse
      Sample Type: Tissue Homogenates
    15. 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;215(2):452-63.
      Species: Human
      Sample Type: Synovial Fluid
    16. 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;105(4):1394-404.
      Species: Mouse
      Sample Type: Tissue Homogenates
    17. 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;106(2):757-69.
      Species: Rat
      Sample Type: Cell Lysates
    18. 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;75(4):1895-903.
      Species: Human
      Sample Type: Cell Culture Supernates
    19. 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;103(17):6735-40.
      Species: Rat
      Sample Type: Whole Tissue
    Expand to show all 19 Citations
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