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Quantikine kits

Manufactured by R&D Systems
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Quantikine kits are a line of quantitative sandwich enzyme-linked immunosorbent assay (ELISA) products developed by R&D Systems for the measurement of specific proteins in cell culture supernatants, serum, plasma, and other biological fluids. These kits provide a standardized and reliable method for the quantitative determination of target analytes.

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Close spelling variants of this product

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80 protocols using quantikine kits

1

Plasma Biomarker Analysis of VEGF, IL-1RA, and IL-8

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Venous blood samples from both HC and CTx patients were collected in the morning using the anticoagulant Citrate Dextrose Solution USP (ACD) Formula A. Two milliliters of plasma were centrifuged (11000 g, 2 min, 4°C) to obtain platelet-free plasma [25 (link)] and the samples were immediately frozen at −80°C. Plasma levels of VEGF-A165 (with no-cross reactivity for other native VEGF-A isoforms or VEGF analogs), IL-1RA and IL-8 were further analyzed by ELISA using the R&D Systems Quantikine kits (DVE00, DRA00B and HS800 respectively; Minneapolis, MN, USA).
Vitiello D., Chaar D., Neagoe P.E., Ducharme A., Carrier M., Pelletier G.B., Racine N., Liszkowski M., Sirois M.G, & White M. (2015). Decreased circulating and neutrophil mediated VEGF-A165 release in stable long-term cardiac transplant recipients. Vascular Cell, 7, 4.
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2

Quantifying Inflammation Biomarkers in PISF

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Total MMP‐8, IL‐6, and calprotectin were measured in the collected PISF samples using ELISA (Quantikine ELISAs, R&D systems). ELISAs were performed according to the manufacturer's protocols. The detection limits of R&D systems Quantikine kits for total MMP‐8, calprotectin, and IL‐6 were 0.013, 0.086, and 0.70 pg/ml, respectively (Lähteenmäki et al., 2020 ).
Lähteenmäki H., Tervahartiala T., Räisänen I.T., Pärnänen P., Mauramo M., Gupta S., Sampson V., Rathnayake N., Heikkinen A., Alassiri S., Gieselmann D., Frankenberger R, & Sorsa T. (2022). Active MMP‐8 point‐of‐care (PoC)/chairside enzyme‐test as an adjunctive tool for early and real‐time diagnosis of peri‐implantitis. Clinical and Experimental Dental Research, 8(2), 485-496.
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3

Cytokine Profile Evaluation in Peritoneal Fluid

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Cytokine levels in peritoneal fluid samples were determined by ELISA using the commercially available human Quantikine kits from R&D Systems (Minneapolis, MN). OPG levels were determined using an ELISA from E Bioscience (Vienna, Austria). The assays were performed in duplicate according to the manufacturer’s protocols. The detection thresholds were 0.79 pg/ml for IL-6, 2.9 pg/ml for IL-10, 7.8 pg/ml for leptin, 4.5 pg/ml for OPG, 33 pg/ml for suPAR and 1.1 ng/ml for CCL18. The intra-assay variability was 5–10 % for IL-6, 2.5–6.6 % for IL-10, 3–3.2 % for leptin, 4.3–7.9 % for OPG, 2.1–7.5 % for suPAR and 3.2–3.7 % for CCL18. The inter-assay variability varied from 3.5 to 7.6 % depending on the cytokine. All samples were examined in duplicate and the median values were used for statistical analysis.
Lane D., Matte I., Garde-Granger P., Laplante C., Carignan A., Rancourt C, & Piché A. (2015). Inflammation-regulating factors in ascites as predictive biomarkers of drug resistance and progression-free survival in serous epithelial ovarian cancers. BMC Cancer, 15, 492.
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4

Apoptosis and Inflammatory Markers Quantification

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Cell apoptotic marker (M30; CK18 fragment), growth factors thrombopoietin (TPO) and chemokines C-X-C motif ligand 5 (ENA-78, CXCL5), C-C motif ligand 2 (MCP-1, CCL2), macrophage inflammatory protein-1a (MIP-1a, CCL3), macrophage inflammatory protein-1b (MIP-1b, CCL4) and regulated upon activation normal T cell expressed and secreted (RANTES, CCL5) were measured by ELISA using the Quantikine Kits (R&D Systems, Wiesbaden, Germany).
Sadeghi M., Lahdou I., Oweira H., Daniel V., Terness P., Schmidt J., Weiss K.H., Longerich T., Schemmer P., Opelz G, & Mehrabi A. (2015). Serum levels of chemokines CCL4 and CCL5 in cirrhotic patients indicate the presence of hepatocellular carcinoma. British Journal of Cancer, 113(5), 756-762.
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5

Cytokine Profiling of Endothelial Cells

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A total of 106 sorted and unsorted EC were grown in EGM-2 for 24 h on 10 cm dishes. After that, the medium was removed, the cells were washed with PBS three times, and unsupplemented basal EBM was added for 120 h to obtain conditioned medium. Cytokine pattern (vascular endothelial growth factor, VEGF; stromal cell-derived factor 1α, SDF1α; hepatocyte growth factor, HGF; epidermal growth factor, EGF; fibroblast growth factor 2, FGF2) was analysed by enzyme-linked immunosorbent assay (ELISA) using Quantikine kits (R&D Systems, USA).
Zakharova I.S., Zhiven’ M.K., Saaya S.B., Shevchenko A.I., Smirnova A.M., Strunov A., Karpenko A.A., Pokushalov E.A., Ivanova L.N., Makarevich P.I., Parfyonova Y.V., Aboian E, & Zakian S.M. (2017). Endothelial and smooth muscle cells derived from human cardiac explants demonstrate angiogenic potential and suitable for design of cell-containing vascular grafts. Journal of Translational Medicine, 15, 54.
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6

Urinary Biomarker Quantification by ELISA

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ELISAs (Quantikine kits; R&D Systems, Inc., Minneapolis, MN, USA) were used to quantify levels of uMMP-2, uMMP-9, uTIMP-1 and uTIMP-2 as previously reported (Smith et al, 2007 (link), 2008 (link)). Specimens, standards and reagents were prepared according to the manufacturer's instructions. Protein concentration in urine was determined by the Bradford method using bovine serum albumin as the standard as previously described by us (Moses et al, 1998 (link); Roy et al, 2004 (link); Pories et al, 2008 (link); Roy et al, 2008 (link)).
Roy R., Zurakowski D., Wischhusen J., Frauenhoffer C., Hooshmand S., Kulke M, & Moses M.A. (2014). Urinary TIMP-1 and MMP-2 levels detect the presence of pancreatic malignancies. British Journal of Cancer, 111(9), 1772-1779.
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7

Adipose Tissue Cytokine Secretion

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Freshly isolated gonadal adipose tissue (AT) was isolated from CD, SD, HF, and HFSD dams at E18. Adipose explants were placed in 24‐well plates (100 mg tissue/well) with 1 mL of complete media (Dulbecco's modified media (DMEM), 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin) for 24 h. Media were harvested and cytokine secretion (TNFα, IL‐6, and IL‐1β) was analyzed by ELISA (Quantikine kits; R&D Systems Europe).
Reynolds C.M., Vickers M.H., Harrison C.J., Segovia S.A, & Gray C. (2014). High fat and/or high salt intake during pregnancy alters maternal meta‐inflammation and offspring growth and metabolic profiles. Physiological Reports, 2(8), e12110.
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8

Imaging and Biomarker Analysis in ICH

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In the FAST trial, baseline labs and biomarkers were collected at the baseline study visit, within 4 hours of ICH symptom onset. Blood samples for cytokine analysis were centrifuged at 3000g for 5 minutes on site, frozen and stored at −80 °C, and analyzed in batches at a central core lab facility. IL-6 was measured with commercially available quantitative sandwich enzyme-linked immunosorbent assay (Quantikine®) kits obtained from R&D Systems, Inc. Laboratory determinations were performed blinded to clinical and neuroimaging findings. Computed tomography (CT) scans were collected at admission and 24 hours after initiation of study treatment. Scans were sent to a central imaging laboratory (Bio-Imaging Technologies) and analyzed in random order with the use of Analyze software (Mayo Clinic) by two neuroradiologists who were blinded to the treatment assignments. As per the FAST protocol, ICH, intraventricular hemorrhage (IVH), and PHE volumes were calculated using a semi-automated process by tracing the perimeter of appropriate high- and low-attenuation zones and calculating lesion areas for each slice multiplied by slice thickness to yield lesion volumes. PHE volumes were calculated by subtracting ICH volume from the combined ICH plus edema volume.20 (link)–22 (link)
Leasure A.C., Kuohn L.R., Vanent K.N., Bevers M.B., Kimberly W.T., Steiner T., Mayer S.A., Matouk C.C., Sansing L.H., Falcone G.J, & Sheth K.N. (2021). Association of Serum IL-6 with Functional Outcome after Intracerebral Hemorrhage. Stroke, 52(5), 1733-1740.
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9

Quantitation of IL-17 in Th17 Cells

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IL-17 production by Th17 polarized cells purified from MLN was measured by solid phase sandwich ELISA using Quantikine kits (R&D systems Inc, MN, USA), according to the manufacturer's instructions: reference number M1700 for IL-17 detection. The assay's sensitivity was 5 pg/ml for IL-17.
Mestre L., Carrillo-Salinas F.J., Mecha M., Feliú A., Espejo C., Álvarez-Cermeño J.C., Villar L.M, & Guaza C. (2019). Manipulation of Gut Microbiota Influences Immune Responses, Axon Preservation, and Motor Disability in a Model of Progressive Multiple Sclerosis. Frontiers in Immunology, 10, 1374.
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10

Cytokine Quantification Protocol

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All cytokines were measured using quantitative sandwich enzyme assay kits (Quantikine kits, R&D Systems, Minneapolis, MN). Sensitivities for cytokines were IL-6 <0.7 pg/ml and IFN-γ <3 pg/ml. The coefficient of variation ranged from 2.6 to 4.9%.
Saban K.L., Collins E.G., Mathews H.L., Bryant F.B., Tell D., Gonzalez B., Bhoopalam S., Chroniak C.P, & Janusek L.W. (2022). Impact of a Mindfulness-Based Stress Reduction Program on Psychological Well-Being, Cortisol, and Inflammation in Women Veterans. Journal of General Internal Medicine, 37(Suppl 3), 751-761.
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