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Human sCD14 Quantikine ELISA Kit

23 citations
  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5 hours
  • Sensitivity
    125 pg/mL
  • Assay Range
    250.00 - 16,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma,)
  • Specificity
    Natural and recombinant human sCD14
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Control Available
Product Summary
The Quantikine Human soluble CD14 Immunoassay is a 4.5 hour solid phase ELISA designed to measure human soluble CD14 in cell culture supernates, serum, and plasma. It contains recombinant human CD14 expressed from CHO cells and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained measuring natural human sCD14 showed dose-response curves that were parallel to the standard curves obtained using the kit standards. These results indicate that this kit can be used to determine relative mass values for natural human sCD14.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Inter-Assay Precision (Precision between assays Three samples of known concentration were tested on one plate to assess intra-assay precision.
Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404040
Mean111121584187115922984346
Standard Deviation7110421686153209
CV%6.44.85.27.46.74.8

Recovery

The recovery of sCD14 spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=5) 91 85-105
Citrate Plasma (n=5) 102 88-110
EDTA Plasma (n=5) 98 90-107
Heparin Plasma (n=5) 96 91-104
Serum (n=5) 98 84-107
Linearity
To assess the linearity of the assay, five samples containing or spiked with high concentrations of sCD14 were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
CD14_DC140_ELISA_42.jpg
Background:

CD14 is a 55 kDa GPI-linked cell surface glycoprotein that is preferentially expressed on monocytes/macrophages. The amino acid sequence of mature human CD14 is approximately 67% and 64% identical to the mouse and rat proteins, respectively.

    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. 100 µL Assay Diluent
    4.   Add 100 µL of Assay Diluent to each well.

    5. 100 µL Standard, Control, or Sample
    6.   Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 3 hours.
    7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

    8. 200 µL Conjugate
    9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 1 hour.
    10.   Aspirate and wash 4 times.

    11. 200 µL Substrate Solution
    12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

    13. 50 µL Stop Solution
    14.   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.

    Showing Results 1 - 10 of 23
    Filter your results:

    Species
    1. Dynamics of immune reconstitution and activation markers in HIV+ treatment-naive patients treated with raltegravir, tenofovir disoproxil fumarate and emtricitabine.
      Authors: Funderburg N, Andrade A, Chan E, Rosenkranz S, Lu D, Clagett B, Pilch-Cooper H, Rodriguez B, Feinberg J, Daar E, Mellors J, Kuritzkes D, Jacobson J, Lederman M
      PLoS ONE, 2013;8(12):e83514.
      Species: Human
      Sample Type: Plasma
    2. SIVagm infection in wild African green monkeys from South Africa: epidemiology, natural history, and evolutionary considerations.
      Authors: Ma, Dongzhu, Jasinska, Anna, Kristoff, Jan, Grobler, J Paul, Turner, Trudy, Jung, Yoon, Schmitt, Christop, Raehtz, Kevin, Feyertag, Felix, Martinez Sosa, Natalie, Wijewardana, Viskam, Burke, Donald S, Robertson, David L, Tracy, Russell, Pandrea, Ivona, Freimer, Nelson, Apetrei, Cristian
      PLoS Pathog, 2013;9(1):e1003011.
      Species: Primate - Chlorocebus pygerythrus (African Green Monkey)
      Sample Type: Plasma
    3. Shared monocyte subset phenotypes in HIV-1 infection and in uninfected subjects with acute coronary syndrome.
      Authors: Funderburg, Nicholas, Zidar, David A, Shive, Carey, Lioi, Anthony, Mudd, Joseph, Musselwhite, Laura W, Simon, Daniel I, Costa, Marco A, Rodriguez, Benigno, Sieg, Scott F, Lederman, Michael
      Blood, 2012;120(23):4599-608.
      Species: Human
      Sample Type: Plasma
    4. Therapeutic helminth infection of macaques with idiopathic chronic diarrhea alters the inflammatory signature and mucosal microbiota of the colon.
      Authors: Broadhurst, Mara Jan, Ardeshir, Amir, Kanwar, Bittoo, Mirpuri, Julie, Gundra, Uma Mahe, Leung, Jacqueli, Wiens, Kirsten, Vujkovic-Cvijin, Ivan, Kim, Charlie, Yarovinsky, Felix, Lerche, Nicholas, McCune, Joseph M
      Oncogene, 2012;8(11):e1003000.
      Species: Human
      Sample Type: Plasma
    5. Relationship between microbial translocation and endothelial function in HIV infected patients.
      Authors: Blodget E, Shen C, Aldrovandi G, Rollie A, Gupta S, Stein J, Dube M
      PLoS ONE, 2012;7(8):e42624.
      Species: Human
      Sample Type: Plasma
    6. Altered monocyte activation markers in Tourette's syndrome: a case-control study.
      Authors: Matz J, Krause DL, Dehning S, Riedel M, Gruber R, Schwarz MJ, Muller N
      BMC Psychiatry, 2012;12(0):29.
      Species: Human
      Sample Type: Serum
    7. Epithelial adhesion molecules can inhibit HIV-1-specific CD8+ T-cell functions.
      Authors: Streeck H, Kwon DS, Pyo A, Flanders M, Chevalier MF, Law K, Julg B, Trocha K, Jolin JS, Anahtar MN, Lian J, Toth I, Brumme Z, Chang JJ, Caron T, Rodig SJ, Milner DA, Piechoka-Trocha A, Kaufmann DE, Walker BD, Altfeld M
      Blood, 2011;117(19):5112-22.
      Species: Human
      Sample Type: Plasma
    8. Traditional risk factors and D-dimer predict incident cardiovascular disease events in chronic HIV infection.
      Authors: Ford ES, Greenwald JH, Richterman AG
      AIDS, 2010;24(10):1509-17.
      Species: Human
      Sample Type: Plasma
    9. Maternal country of origin, breast milk characteristics and potential influences on immunity in offspring.
      Authors: Holmlund U, Amoudruz P, Johansson MA, Haileselassie Y, Ongoiba A, Kayentao K, Traore B, Doumbo S, Schollin J, Doumbo O, Montgomery SM, Sverremark-Ekstrom E
      Clin. Exp. Immunol., 2010;162(3):500-9.
      Species: Human
      Sample Type: Milk
    10. Increased tissue factor expression on circulating monocytes in chronic HIV infection: relationship to in vivo coagulation and immune activation.
      Authors: Funderburg NT, Mayne E, Sieg SF
      Blood, 2010;115(2):161-7.
      Species: Human
      Sample Type: Plasma
    11. Cutting edge: Experimentally induced immune activation in natural hosts of simian immunodeficiency virus induces significant increases in viral replication and CD4+ T cell depletion.
      Authors: Pandrea I, Gaufin T, Brenchley JM, Gautam R, Monjure C, Gautam A, Coleman C, Lackner AA, Ribeiro RM, Douek DC, Apetrei C
      J. Immunol., 2008;181(10):6687-91.
      Species: Primate – Chlorocebus pygerythrus (Vervet/African Green Monkey)
      Sample Type: Plasma
    12. Ethnic and sex differences in circulating endotoxin levels: A novel marker of atherosclerotic and cardiovascular risk in a British multi-ethnic population.
      Authors: Miller MA, McTernan PG, Harte AL, Silva NF, Strazzullo P, Alberti KG, Kumar S, Cappuccio FP
      Atherosclerosis, 2009;203(2):494-502.
      Species: Human
      Sample Type: Plasma
    13. Breast milk fatty acids may link innate and adaptive immune regulation: analysis of soluble CD14, prostaglandin E2, and fatty acids.
      Authors: Laitinen K, Hoppu U, Hamalainen M, Linderborg K, Moilanen E, Isolauri E
      Pediatr. Res., 2006;59(5):723-7.
    14. CD14 is an acute-phase protein.
      Authors: Bas S, Gauthier BR, Spenato U, Stingelin S, Gabay C
      J. Immunol., 2004;172(7):4470-9.
    15. Reduced interferon gamma production and soluble CD14 levels in early life predict recurrent wheezing by 1 year of age.
      Authors: Guerra S, Lohman IC, Halonen M, Martinez FD, Wright AL
      Am. J. Respir. Crit. Care Med., 2003;169(1):70-6.
      Species: Human
      Sample Type: Serum
    16. Shift in monocyte apoptosis with increasing viral load and change in apoptosis-related ISG/Bcl2 family gene expression in chronically HIV-1-infected subjects.
      Authors: Patro S, Pal S, Bi Y, Lynn K, Mounzer K, Kostman J, Davuluri R, Montaner L
      J Virol, 2015;89(1):799-810.
      Species: Human
      Sample Type: Plasma
    17. Plasma sCD14 as a biomarker to predict pulmonary exacerbations in cystic fibrosis.
      Authors: Quon B, Ngan D, Wilcox P, Man S, Sin D
      PLoS ONE, 2014;9(2):e89341.
      Species: Human
      Sample Type: Plasma
    18. Transcriptional effects of 1,25 dihydroxyvitamin D(3) physiological and supra-physiological concentrations in breast cancer organotypic culture.
      Authors: Milani C, Katayama M, de Lyra E, Welsh J, Campos L, Brentani M, Maciel M, Roela R, del Valle P, Goes J, Nonogaki S, Tamura R, Folgueira M
      BMC Cancer, 2013;13(0):119.
      Species: Human
      Sample Type: cell culture supernate
    19. CD4 T follicular helper cell dynamics during SIV infection.
      J. Clin. Invest., 2012;122(9):3281-94.
      Species: Primate - Macaca mulatta (Rhesus monkey)
      Sample Type: Plasma
    20. The relationship of systemic inflammation to prior hospitalization in adult patients with cystic fibrosis.
      Authors: Ngan D, Wilcox P, Aldaabil M, Li Y, Leipsic J, Sin D, Man S
      BMC Pulm Med, 2012;12(0):3.
      Species: Human
      Sample Type: Plasma
    21. TGF-beta in human milk is associated with wheeze in infancy.
      Authors: Oddy WH, Halonen M, Martinez FD, Lohman IC, Stern DA, Kurzius-Spencer M, Guerra S, Wright AL
      J. Allergy Clin. Immunol., 2003;112(4):723-8.
      Species: Human
      Sample Type: Milk
    22. Elevated plasma soluble CD14 and skewed CD16+ monocyte distribution persist despite normalisation of soluble CD163 and CXCL10 by effective HIV therapy: a changing paradigm for routine HIV laboratory monitoring?
      Authors: Castley, Alison, Berry, Cassandr, French, Martyn, Fernandez, Sonia, Krueger, Romano, Nolan, David
      PLoS ONE, 2014;9(12):e115226.
      Species: Human
      Sample Type: Plasma
    23. Large number of rebounding/founder HIV variants emerge from multifocal infection in lymphatic tissues after treatment interruption.
      Authors: Rothenberger, Meghan K, Keele, Brandon, Wietgrefe, Stephen, Fletcher, Courtney, Beilman, Gregory, Chipman, Jeffrey, Khoruts, Alexande, Estes, Jacob D, Anderson, Jodi, Callisto, Samuel P, Schmidt, Thomas E, Thorkelson, Ann, Reilly, Cavan, Perkey, Katherin, Reimann, Thomas G, Utay, Netanya, Nganou Makamdop, Krystell, Stevenson, Mario, Douek, Daniel C, Haase, Ashley T, Schacker, Timothy
      Proc Natl Acad Sci U S A, 2015;112(10):E1126-34.
      Species: Human
      Sample Type: Plasma
    Expand to show all

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      Catalog # DC140

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