Human cd14 quantikine elisa kit

Manufactured by R&D Systems

Sourced in United States

The Human CD14 Quantikine ELISA Kit is a quantitative sandwich enzyme immunoassay designed to measure human CD14 levels in cell culture supernates, serum, and plasma. It is a tool for the quantitative determination of human CD14 concentrations in biological fluids.

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Quantikine ELISA kit (1163 citations) Quantikine ELISA (398 citations) ELISAs (144 citations) Quantikine kits (80 citations) Quantikines (4 citations) Human CD14 Quantikine Kit (3 citations) CD14 ELISA kit (2 citations) Quantikine Human (2 citations) Human kits (2 citations) Human soluble CD14 Quantikine ELISA Kit (2 citations)

15 protocols using human cd14 quantikine elisa kit

Plasma Biomarkers of Inflammation

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Levels of soluble CD14 (sCD14), soluble CD163 (sCD163), neopterin, LPS, soluble IFN-γ–induced protein 10 (IP-10), and C-reactive protein (CRP) were quantified in plasma using commercially available ELISA kits according to the manufacturer’s instructions. sCD14 levels were quantified using a human CD14 Quantikine ELISA kit (R&D Systems) and expressed as μg/mL. IP-10 levels were quantified using a human IP-10 Quantikine ELISA kit (R&D Systems) and expressed as pg/ml. Plasma CRP levels were measured using a monkey CRP ELISA kit (Life Diagnostics Inc.) and expressed as μg/ml. sCD163 levels were quantified using a Macro163 ELISA kit (IQ Products and Trillium Diagnostics) and expressed as ng/mL. Neopterin levels were quantified using an ELISA for the quantitative determination of neopterin in serum, plasma, and urine (Brahms Diagnostica) and expressed as ng/mL. For quantification of LPS levels, plasma samples were diluted to 20% with endotoxin-free water and heated to 70°C for 10 minutes followed by quantification using a Limulus Amebocyte assay (Cambrex) expressed as pg/mL.

Harper J., Ribeiro S.P., Chan C.N., Aid M., Deleage C., Micci L., Pino M., Cervasi B., Raghunathan G., Rimmer E., Ayanoglu G., Wu G., Shenvi N., Barnard R.J., Del Prete G.Q., Busman-Sahay K., Silvestri G., Kulpa D.A., Bosinger S.E., Easley K.A., Howell B.J., Gorman D., Hazuda D.J., Estes J.D., Sekaly R.P, & Paiardini M. (2022). Interleukin-10 contributes to reservoir establishment and persistence in SIV-infected macaques treated with antiretroviral therapy. The Journal of Clinical Investigation, 132(8), e155251.

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Plasma Cytokine and sCD14 Quantification

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Plasma samples were analysed using a custom 24-plex magnetic Luminex assay (R&D systems). All samples were diluted 1:2 and assays were performed according to manufacturer’s protocol. Samples were acquired using the Bio-Plex Magpix multiplex reader (Bio-Rad) and analysed using the Bio-Plex Manager software. Soluble CD14 (sCD14) concentration in plasma samples was measured using the human CD14 Quantikine ELISA kit from R&D systems (DC140) according to manufacturer’s protocol.

Gorin J.B., Malone D.F., Strunz B., Carlsson T., Aleman S., Björkström N.K., Falconer K, & Sandberg J.K. (2020). Plasma FABP4 is associated with liver disease recovery during treatment-induced clearance of chronic HCV infection. Scientific Reports, 10, 2081.

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Quantifying Inflammatory and Coagulation Markers

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Serum and soluble inflammatory and coagulation markers were obtained using commercially available kits. Assays were performed according to the manufacture’s protocols. The following markers were measured in blood: pro-inflammatory cytokines serum IL-6 (Human IL-6 Qunatikine HS Elisa Kit), serum IL-18 (Human IL-18/IL-1 F4 ELISA), soluble TNF-α receptor-1 (TNFR1) (Human sTNF RI/TNFRSF1A Quantikine ELISA Kit), and soluble TNF-α receptor-2 (TNFR2) (Human sTNF RII/TNFRSF1B Quantikine ELISA Kit), Interferon gamma-induced protein-10 (IP-10)(Human CXCL10/IP-10 Quantikine ELISA Kit), soluble CD14 (Human CD14 Quantikine ELISA Kit) (all R&D System, Minneapolis, MN, USA) and pro-inflammatory protein C-reactive protein (CRP) (EIA kit, Cayman Chemical Company, Ann Arbor, MI, USA), Serum Amyloid A (SAA) (ELISA, Assaypro, St. Charles, MO, USA) and were also measured. Inflammatory index is a calculated parameter: 0.333*log (IL-6) + 0.666*log (sTNFR1). The following markers of thrombosis and coagulation were also measured: Plasminogen activator inhibitor-1 (PAI-1) (AssayMax Human PAI-1 ELISA kit, Assaypro, St. Charles, MO, USA), D-dimer (ELISA ZYMUTEST DDimer, ANIARA, West Chester, OH, USA) and Fibrinogen (immunoperoxidase assay, GenWay, San Diego, CA, USA).

Van Epps P., Oswald D., Higgins P.A., Hornick T.R., Aung H., Banks R.E., Wilson B.M., Burant C., Graventstein S, & Canaday D.H. (2016). Frailty has a stronger association with inflammation than age in older veterans. Immunity & Ageing : I & A, 13, 27.

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Plasma Biomarkers of Coagulation, Inflammation, and Bacterial Translocation

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We measured levels of D-dimer, IL6, hs-CRP, IFABP, sCD14, and sCD163 in plasma as indicators of coagulation, inflammation, monocyte activation, and bacterial translocation using the commercial ELISA assays Human D2D (D-Dimer) ELISA Kit (ImmunoWay Biotechnology, Plano, Texas), Human Quantikine IL-6 HS ELISA (R&D Systems, Minneapolis, Minnesota), CRP ELISA (LDN Labor Diagnostika Nord, Nordhorn Germany), PicoKine ELISA for Human IFABP (Boster Bio, Pleasanton, California), Human CD14 Quantikine ELISA Kit (R&D Systems) and Human CD163 Quantikine ELISA Kit (R&D Systems), respectively, accordingly to manufacturers’ instructions. Each sample was assayed in duplicate in batches seeking an equal representation of the study groups and containing all samples from the same individual. Data was collected using a microplate luminometer with Gen5 Data Analysis Software (Biotek, Winooski, Vermont).

Serrano-Villar S., López-Huertas M.R., Jiménez D., Galera C., Martínez-Sanz J., Moreno E., Muriel A., Gutiérrez F., Busca C., Portilla J., Bisbal O., Iribarren J.A., Tejerina F., de los Santos I, & Moreno S. (2022). Long-Term Changes of Inflammatory Biomarkers in Individuals on Suppressive Three-Drug or Two-Drug Antiretroviral Regimens. Frontiers in Immunology, 13, 848630.

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Plasma Biomarker Analysis in IgAN

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Plasma was obtained from whole blood, drawn in EDTA tube, by centrifuging at 400xg, for 10 min at room temperature. Plasma samples from IgAN patients and HC were then analyzed with enzyme-linked immunosorbent assay (ELISA) to determine the concentration of the following factors; monocyte chemoattractant protein (MCP-1) (Human CCL2/MCP-1 Quantikine ELISA Kit, R&D Systems, USA), soluble CD14 (Human CD14 Quantikine ELISA Kit, R&D Systems), soluble CD40L (Human CD40 Ligand/TNFSF5 Quantikine ELISA Kit, R&D Systems), B cell activating factor (BAFF) (Human BAFF Quantikine ELISA Kit, R&D Systems), IL-6 (Human IL-6 Quantikine HS ELISA Kit, R&D Systems), Fractalkine (Human CX3CL1/Fractalkine Quantikine ELISA Kit, R&D Systems) and MIP-1 (Human CCL3/MIP-1 alpha Quantikine ELISA Kit, R&D Systems). Catalog number for ELISA kits are shown in S2 Table. The samples were assayed in duplicates and the average values were considered for further analysis. The optical density (OD) in blank well was subtracted from the OD values in samples before calculation of concentrations.

Sendic S., Mansouri L., Lundberg S., Nopp A., Jacobson S.H, & Lundahl J. (2021). B cell and monocyte phenotyping: A quick asset to investigate the immune status in patients with IgA nephropathy. PLoS ONE, 16(3), e0248056.

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Cytokine and sCD14 Quantification

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Supernatants from GM-MØ were tested for the presence of IL-6, IL-10, TNFα, and CXCL10 (BioLegend) and IFNβ (R&D Systems). For the detection of sCD14 in supernatants, serum, and plasma of RA patients and healthy controls, the Human CD14 Quantikine ELISA Kit (R&D Systems) was used.

Fuentelsaz-Romero S., Barrio-Alonso C., García Campos R., Torres Torresano M., Muller I.B., Triguero-Martínez A., Nuño L., Villalba A., García-Vicuña R., Jansen G., Miranda-Carús M.E., González-Álvaro I, & Puig-Kröger A. (2021). The Macrophage Reprogramming Ability of Antifolates Reveals Soluble CD14 as a Potential Biomarker for Methotrexate Response in Rheumatoid Arthritis. Frontiers in Immunology, 12, 776879.

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Multiplex Biomarker Measurement in Plasma

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Cryopreserved plasma aliquots were thawed and prepared following kit manufacturer’s guidelines. MCP-1, sCD163, and IP-10 were measured using a custom multiplex kit (RD Systems, MN, USA), acquired on a Luminex 200™ analyzer (Luminex), and analyzed using Bio-Plex Manager™ software (Bio-Rad). Neopterin and sCD14 were measured by ELISA (Neopterin competitive enzyme immunoassay, ALPCO, NH, USA; Human CD14 Quantikine ELISA kit, RD Systems), optical density read on a microplate spectrophotometer (Bio-Rad), and standard curve interpolation conducted on Prism version 7.0b (Graphpad Software Inc., CA, USA). Samples were run in duplicate and the CV of replicates were less than 10%.
Examination of biomarker distributions revealed two outliers greater than 4 SDs above the biomarker distribution means – one for neopterin and one for MCP-1. Because these values were considered physiologically plausible, we substituted these outlier values with the next most extreme value in the biomarker distribution during statistical analysis, as has been done previously by our group [15 (link)]. The biomarker distributions of sCD163 and IP-10 were log₁₀ transformed prior to statistical analysis.

CHANG K., PREMEAUX T.A., COBIGO Y., MILANINI B., HELLMUTH J., RUBIN L.H., JAVANDEL S., ALLEN I., NDHLOVU L.C., PAUL R, & VALCOUR V. (2020). Plasma Inflammatory Biomarkers Link to Diffusion Tensor Imaging Metrics in Virally Suppressed HIV-Infected Individuals. AIDS (London, England), 34(2), 203-213.

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Multiplex Inflammatory Biomarkers Assay

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Quantification of inflammatory cytokines (IL12, TNF, IL10, IL6, IL1b, IL8) was accomplished using BD Cytometric Bead Array Human Inflammatory Cytokines Kit (BD BioScience). Soluble CD14 was measured using Human CD14 Quantikine ELISA Kit (R&D Systems, BioTechne). Intestinal fatty acid-binding protein (I-FABP) was measured using Human FABP2/I-FABP Quantikine ELISA Kit (R&D Systems, BioTechne). Lipopolysaccharide-binding protein (LBP) was measured using Human LBP ELISA kit (ThermoFisher). Endotoxin was measured using Pierce™ Chromogenic Endotoxin Quant Kit (ThermoFisher). All procedures were performed using plasma samples according to manufacturers’ protocols.

Jamieson P.E., Smart E.B., Bouranis J.A., Choi J., Danczak R.E., Wong C.P., Paraiso I.L., Maier C.S., Ho E., Sharpton T.J., Metz T.O., Bradley R, & Stevens J.F. (2024). Gut enterotype-dependent modulation of gut microbiota and their metabolism in response to xanthohumol supplementation in healthy adults. Gut Microbes, 16(1), 2315633.

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Quantification of Milk Biomarkers

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OPN, TGF-β1, and sCD14 were measured using a Human Osteopontin Quantikine ELISA Kit, a Human TGF-beta 1 Quantikine ELISA Kit, and a Human CD14 Quantikine ELISA Kit, respectively (R&D Systems Inc, Minneapolis, MN, USA). Before the analysis, the milk samples were thawed and centrifuged to remove residual cream or precipitate as per the assay manufacturer’s instructions [6 (link),20 (link),21 (link)] and as described previously [20 (link),21 (link),23 (link)]. The cytokine levels were converted to concentrations using a five-parameter logistic model. The LLOQs for OPN, TGF-β1, and sCD14 were 0.31 ng/mL, 31.3 pg/mL, and 250 pg/mL, respectively. Cytokines below the LLOQ were judged to be negative and were allocated the concentration of the LLOQ to aid the statistical analysis.

Takahashi T., Ueno H.M., Yamaide F., Nakano T., Shiko Y., Kawasaki Y., Mitsuishi C, & Shimojo N. (2023). Comparison of 30 Cytokines in Human Breast Milk between 1989 and 2013 in Japan. Nutrients, 15(7), 1735.

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Comprehensive Cytokine and Biomarker Profiling

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Plasma aliquots were thawed and prepared following kit manufacturer guidelines. All samples were analyzed in duplicate. MCP-1, soluble (s)CD163, IP-10, galectin-1, galectin-3, galectin-9, IL-6, TNF-α, TNFR1, D-dimer, and cystatin C were measured using custom Luminex kits from (R&D Systems) and CRP was measured using the Procartaplex kit (Thermofisher). Data was acquired on a Luminex 200^TM analyzer (Luminex) and analyzed using MILLIPLEX^® Analyst software (Millipore). Neopterin and sCD14 were measured via ELISA (Neopterin competitive enzyme immunoassay, ALPCO, NH, USA; Human CD14 Quantikine ELISA kit, R&D Systems). Optical density was read with a microplate spectrophotometer (Bio-Rad) and data analysis, including four parameter logistic standard interpolation, was carried out using the online MyAssays Ltd. data analysis tool.

Premeaux T.A., Javandel S., Hosaka K.R., Greene M., Therrien N., Allen I.E., Corley M.J., Valcour V.G, & Ndhlovu L.C. (2020). Associations Between Plasma Immunomodulatory and Inflammatory Mediators With VACS Index Scores Among Older HIV-Infected Adults on Antiretroviral Therapy. Frontiers in Immunology, 11, 1321.

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