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Human CD14 DuoSet ELISA

Catalog #: DY383
25 Citations 2 Reviews Datasheet / COA / SDS

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DY383 has been discontinued.
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Human CD14 ELISA Standard Curve
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Citations (25)
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Summary Product Features Kit Content Other Reagents Required Data Examples Product Datasheets Preparation and Storage Background

Human CD14 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
62.5 - 4,000 pg/mL
Sufficient Materials
For five or fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human CD14. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Scientific Data

N/A Human CD14 ELISA Standard Curve View Larger

Human CD14 ELISA Standard Curve

Product Datasheets

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Product Datasheet
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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: CD14

CD14 is a glycoprotein that mediates the interaction of lipopolysaccharide (LPS, endotoxin) with cells, thereby signaling the presence of gram-negative bacteria (1-3). CD14 is either soluble (CD14) (4, 5) or membrane-bound (mCD14) by a glycosylphosphatidylinositol (GPI) anchor (6, 7). mCD14 is a 55 kDa glycoprotein (1), while CD14 varies from about 43 to 53 kDa, depending on the degree of glycosylation and whether it was synthesized without the anchor or was shed by phospholipase cleavage of the anchor or by proteolysis (12-14). There is no evidence for different mRNAs for m- and CD14. There is no apparent sequence homology with other proteins. The sequence of human CD14 is 63-73% identical to that of mouse, rat, or rabbit CD14 (15). 

mCD14 is expressed primarily on myeloid cells, such as monocytes, macrophages and neutrophils (1-3), the cells most sensitive to LPS, and to a lesser extent on other cells, such as B cells (8) and a circulating dendritic cell progenitor (9). CD14 appears to mediate LPS stimulation of cells that do not express mCD14 (10, 11), such as endothelial, epithelial and smooth-muscle cells. CD14 is found in both serum and urine (5).  
The binding of LPS to CD14 requires an acute phase protein, LPS-binding protein (LBP) (16). The relationship of mCD14, CD14, LPS and LBP is complicated. At low concentrations of LPS, LBP is essential for the binding of LPS to CD14, but at high concentrations, LBP may actually inhibit binding of LPS to CD14. In addition, CD14 may compete with mCD14 for LPS (17) and may serve to help clear LPS (18). These four factors thus appear to participate in a complex feedback mechanism of immune regulation involving both up-regulation and down-regulation of the inflammatory process triggered by LPS. It is loss of control of this mechanism that appears to lead to septic shock. LPS-bound CD14 signals production of inflammatory cytokines and other inflammatory proteins, but the mechanism of signal transduction is unclear. Since a GPI anchor is not transmembrane, there presumably is another transmembrane protein on cells through which LPS-bound CD14 transmits a signal (19). 
In addition to its well known role in gram-negative infections, CD14 likely serves other functions as well. It recognizes soluble peptidylglycan from gram-positive cell walls (20), and it has been reported to bind apoptotic cells and induce their phagocytosis (21).

Entrez Gene IDs:
929 (Human); 12475 (Mouse); 60350 (Rat); 100037938 (Porcine)
Alternate Names:
CD14 antigen; CD14 molecule; CD14; monocyte differentiation antigen CD14; Myeloid cell-specific leucine-rich glycoprotein

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Human CD14 DuoSet ELISA

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.

25 Citations: Showing 1 - 10
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  1. Intestinal Barrier Dysfunction and Microbial Translocation in Patients with First-Diagnosed Atrial Fibrillation
    Authors: L Blöbaum, M Witkowski, M Wegner, S Lammel, PA Schencke, K Jakobs, M Puccini, D Rei beta ner, D Steffens, U Landmesser, U Rauch, J Friebel
    Biomedicines, 2023-01-10;11(1):.
    Species: Human
    Sample Types: Plasma
  2. Intestinal Injury Biomarkers Predict Mortality in Pediatric Severe Malaria
    Authors: ML Sarangam, R Namazzi, D Datta, C Bond, CPB Vanderpool, RO Opoka, CC John, AL Conroy
    MBio, 2022-09-07;0(0):e0132522.
    Species: Human
    Sample Types: Serum
  3. Immune responses in Omicron SARS-CoV-2 breakthrough infection in vaccinated adults
    Authors: H Kared, AS Wolf, A Alirezayla, A Ravussin, G Solum, TT Tran, F Lund-Johan, JT Vaage, LS Nissen-Mey, UC Nygaard, O Hungnes, AH Robertson, LM Næss, L Trogstad, P Magnus, LA Munthe, S Mjaaland
    Nature Communications, 2022-07-18;13(1):4165.
    Species: Human
    Sample Types: Plasma
  4. Irreversible depletion of intestinal CD4+ T-cells is associated with T-cell activation during chronic HIV infection
    Authors: OE Asowata, A Singh, A Ngoepe, N Herbert, R Fardoos, K Reddy, Y Zungu, F Nene, N Mthabela, D Ramjit, F Karim, K Govender, T Ndung'u, JZ Porterfiel, JH Adamson, FG Madela, VT Manzini, F Anderson, A Leslie, HN Kløverpris
    JCI Insight, 2021-11-22;0(0):.
    Species: Human
    Sample Types: Plasma
  5. Complement activation is associated with poor outcome after out-of-hospital cardiac arrest
    Authors: V Chaban, ER Nakstad, H Stær-Jense, C Schjalm, I Seljeflot, J Vaage, C Lundqvist, JŠ Benth, K Sunde, TE Mollnes, GØ Andersen, SE Pischke
    Resuscitation, 2021-06-11;0(0):.
    Species: Human
    Sample Types: Plasma
  6. Changes in liver steatosis in HIV-positive women are associated with the BMI, but not with biomarkers
    Authors: R Fernandez-, MW Plankey, D Ware, J Bordon
    Cytokine, 2021-05-12;0(0):155573.
    Species: Human
    Sample Types: Plasma
  7. Exploring the potential effect of paricalcitol on markers of inflammation in de novo renal transplant recipients
    Authors: HK Pihlstrøm, T Ueland, AE Michelsen, P Aukrust, F Gatti, C Hammarströ, M Kasprzycka, J Wang, G Haraldsen, G Mjøen, DO Dahle, K Midtvedt, IA Eide, A Hartmann, H Holdaas
    PLoS ONE, 2020-12-16;15(12):e0243759.
    Species: Human
    Sample Types: Plasma
  8. Persistence of monocyte activation under treatment in people followed since acute HIV-1 infection relative to participants at high or low risk of HIV infection
    Authors: S Novelli, C Lécuroux, C Goujard, J Reynes, A Villemant, L Blum, A Essat, V Avettand-F, O Launay, JM Molina, C Bourgeois, L Meyer
    EBioMedicine, 2020-11-26;62(0):103129.
    Species: Human
    Sample Types: Plasma
  9. Expression of Matrix Metalloproteinases and Their Tissue Inhibitors in Peripheral Blood Leukocytes and Plasma of Children with Nonalcoholic Fatty Liver Disease
    Authors: JB Trojanek, J Micha?kiew, R Grzywa-Czu, W Ja?czyk, L Gackowska, I Kubiszewsk, A Helmin-Bas, A Wierzbicka, M Szalecki, P Socha
    Mediators Inflamm., 2020-09-10;2020(0):8327945.
    Species: Human
    Sample Types: Plasma
  10. Urinary proteomics links keratan sulfate degradation and lysosomal enzymes to early type 1 diabetes
    Authors: JAD Van, S Clotet-Fre, AC Hauschild, I Batruch, I Jurisica, Y Elia, FH Mahmud, E Sochett, EP Diamandis, JW Scholey, A Konvalinka
    PLoS ONE, 2020-05-26;15(5):e0233639.
    Species: Human
    Sample Types: Urine
  11. HIV-associated gut dysbiosis is independent of sexual practice and correlates with noncommunicable diseases
    Authors: I Vujkovic-C, O Sortino, E Verheij, J Sklar, FW Wit, NA Kootstra, B Sellers, JM Brenchley, J Ananworani, MSV Loeff, Y Belkaid, P Reiss, I Sereti
    Nat Commun, 2020-05-15;11(1):2448.
    Species: Human
    Sample Types: Plasma
  12. Association between the cytokine storm, immune cell dynamics, and viral replicative capacity in hyperacute HIV infection
    Authors: DM Muema, NA Akilimali, OC Ndumnego, SS Rasehlo, R Durgiah, DBA Ojwach, N Ismail, M Dong, A Moodley, KL Dong, ZM Ndhlovu, JM Mabuka, BD Walker, JK Mann, T Ndung'u
    BMC Med, 2020-03-25;18(1):81.
    Species: Human
    Sample Types: Plasma
  13. Inflammatory Bowel Disease Types Differ in Markers of Inflammation, Gut Barrier and in Specific Anti-Bacterial Response
    Authors: S Coufal, N Galanova, L Bajer, Z Gajdarova, D Schierova, Z Jiraskova, K Kostovciko, Z Jackova, Z Stehlikova, P Drastich, H Tlaskalova, M Kverka
    Cells, 2019-07-13;8(7):.
    Species: Human
    Sample Types: Serum
  14. The effects of CD14 and IL-27 on induction of endotoxin tolerance in human monocytes and macrophages
    Authors: C Petes, V Mintsopoul, RL Finnen, BW Banfield, K Gee
    J. Biol. Chem., 2018-09-21;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  15. Soluble CD14 as a Diagnostic Biomarker for Smear-Negative HIV-Associated Tuberculosis
    Authors: Y Liu, OC Ndumnego, T Chen, RS Kim, ER Jenny-Avit, T Ndung'u, D Wilson, JM Achkar
    Pathogens, 2018-02-27;7(1):.
    Species: Human
    Sample Types: Serum
  16. Impact of early cART on HIV blood and semen compartments at the time of primary infection
    Authors: A Chéret, C Durier, A Mélard, M Ploquin, J Heitzmann, C Lécuroux, V Avettand-F, L David, G Pialoux, JM Chennebaul, M Müller-Tru, C Goujard, C Rouzioux, L Meyer
    PLoS ONE, 2017-07-14;12(7):e0180191.
    Species: Human
    Sample Types: Plasma
  17. Monocyte/macrophage and T cell activation markers are not independently associated with MI risk in healthy individuals - results from the HUNT Study
    Authors: T Ueland, LE Laugsand, LJ Vatten, I Janszky, C Platou, AE Michelsen, JK Damås, P Aukrust, BO Åsvold
    Int. J. Cardiol., 2017-06-12;0(0):.
    Species: Human
    Sample Types: Serum
  18. High Cellular Monocyte Activation in People Living With Human Immunodeficiency Virus on Combination Antiretroviral Therapy and Lifestyle-Matched Controls Is Associated With Greater Inflammation in Cerebrospinal Fluid
    Authors: T Booiman, FW Wit, I Maurer, D De Frances, CA Sabin, AM Harskamp, M Prins, P Garagnani, C Pirazzini, C Franceschi, D Fuchs, M Gisslén, A Winston, P Reiss, NA Kootstra
    Open Forum Infect Dis, 2017-05-25;4(3):ofx108.
    Species: Human
    Sample Types: Plasma
  19. Identification of promising plasma immune biomarkers to differentiate active pulmonary tuberculosis
    Cytokine, 2016-08-31;88(0):99-107.
    Species: Human
    Sample Types: Plasma
  20. Intestinal cell damage and systemic immune activation in individuals reporting sensitivity to wheat in the absence of coeliac disease.
    Authors: Uhde M, Ajamian M, Caio G, De Giorgio R, Indart A, Green P, Verna E, Volta U, Alaedini A
    Gut, 2016-07-25;65(12):1930-1937.
    Species: Human
    Sample Types: Serum
  21. Associations between Cognition, Gender and Monocyte Activation among HIV Infected Individuals in Nigeria.
    Authors: Royal W, Cherner M, Burdo T, Umlauf A, Letendre S, Jumare J, Abimiku A, Alabi P, Alkali N, Bwala S, Okwuasaba K, Eyzaguirre L, Akolo C, Guo M, Williams K, Blattner W
    PLoS ONE, 2016-02-01;11(2):e0147182.
    Species: Human
    Sample Types: Plasma
  22. 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, 2014-02-20;9(2):e89341.
    Species: Human
    Sample Types: Plasma
  23. Daily exposure to dust alters innate immunity.
    Authors: Sahlander K, Larsson K, Palmberg L
    PLoS ONE, 2012-02-15;7(2):e31646.
    Species: Human
    Sample Types: Serum
  24. Integrated proteomic analysis of human cancer cells and plasma from tumor bearing mice for ovarian cancer biomarker discovery.
    Authors: Pitteri SJ, JeBailey L, Faca VM, Thorpe JD, Silva MA, Ireton RC, Horton MB, Wang H, Pruitt LC, Zhang Q, Cheng KH, Urban N, Hanash SM, Dinulescu DM
    PLoS ONE, 2009-11-19;4(11):e7916.
    Species: Human
    Sample Types: Plasma
  25. Soluble CD14 and toll-like receptor-2 are potential salivary biomarkers for oral lichen planus and burning mouth syndrome.
    Authors: Srinivasan M, Kodumudi KN, Zunt SL
    Clin. Immunol., 2007-10-03;126(1):31-7.
    Species: Human
    Sample Types: Saliva

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Human CD14 DuoSet ELISA
By Anonymous on 04/02/2020
Sample Tested: Serum and Plasma

We used this kit for the quantification of CD14 in human serum and plasma. Works very well and well-described protocol.

Human CD14 DuoSet ELISA DY383
Human CD14 DuoSet ELISA
By Anonymous on 10/04/2019
Sample Tested: Serum and Plasma
Human CD14 DuoSet ELISA DY383