Human quantikine kits

Manufactured by R&D Systems

Sourced in United States

The Human Quantikine kits are a series of quantitative sandwich immunoassay kits designed to measure the concentrations of various human proteins in cell culture supernates, serum, plasma, and other biological fluids. These kits utilize a microplate format and employ the quantitative sandwich enzyme immunoassay technique.

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Lab products found in correlation

Lal qcl 1000 assay, by Lonza (1 mentions) Hk315, by Hycult Biotech (1 mentions)

Close spelling variants of this product

Quantikine ELISA human kits Human Quantikine® Immunoassay kits Human TGF-β2 Quantikine ELISA Kits Quantikine human insulin and LPa assay kits Quantikine high sensitivity human IL-6 ELISA kits Quantikine ELISA Human Amyloid β aa1-40/aa1-42 immunoassay kits Quantikine High Sensitivity human IL-6 kits Quantikine kits Quantikine Human Human kits

6 protocols using human quantikine kits

Quantification of Inflammatory Markers in Liver Disease

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To measure BT-related markers and proinflammatory cytokines, peripheral blood samples were collected on the same date of pre- and post-treatment HVPG measurements. Blood samples were separated by centrifugation at 1,500×g for 15 minutes within 30 minutes of collection. Samples were stored at −80°C until the analysis.
To determine LPS, serum samples were diluted 1:10 with pyrogen-free water and incubated for 10 minutes at 75°C to remove serum inhibitors. The concentration of LPS in serum was analyzed using the Limulus Amebocyte Lysate (LAL) assay, a quantitative chromogenic test for detecting endotoxins (QCL-1000 LAL assay; Lonza, Walkersville, MD, USA). The lower detection limit for LPS was 0.1 EU/mL. ELISA assays were used to quantitatively measure the serum concentration of LBP (HK315; Hycult Biotech, Uden, The Netherlands), IL-6 (Human Quantikine kits; R&D Systems, Minneapolis, MN, USA), and TNF-α (Human Quantikine kits) according to the manufacturers’ instructions. The absorbance at the 450-nm light wave was measured in each well with a microplate reader (BioTek ELX; BioTek, Shoreline, WA, USA). The lower limit of detection for LBP was 4.4 ng/mL, IL-6 was 0.70 pg/mL, and TNF-α was 0.5 pg/mL. All measurements were performed in duplicate and the mean value is presented.

Lim Y.L., Kim M.Y., Jang Y.O., Baik S.K, & Kwon S.O. (2017). Rifaximin and Propranolol Combination Therapy Is More Effective than Propranolol Monotherapy for the Reduction of Portal Pressure: An Open Randomized Controlled Pilot Study. Gut and Liver, 11(5), 702-710.

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Quantifying Cytokine and Nitric Oxide Levels

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Enzyme-linked immunosorbent assays (ELISAs) for the quantitative measurement of TNF-α and IL-6 levels were carried out in serum samples of patients by handling Human Quantikine kits (R&D Systems, Minneapolis, MN), according to manufacturer’s instructions. All samples were tested in triplicate and read in a microplate reader. Lower limits of detection of all cytokine assays were 5–8 pg/mL. Standard curves were generated for each plate, and the average zero standard optical densities were subtracted from the rest of standards, controls, and samples to obtain a corrected concentration for both cytokines.
The sum of the NO metabolites nitrite (NO₂⁻) and nitrate (NO₃⁻) is widely used as an index of NO generation32 (link) and expressed as NOx levels (nmol/ml). NOx levels were calculated by measuring conversion of NO₃⁻ to NO₂⁻ by the enzyme nitrate reductase using an ELISA assay (R&D Systems, Minneapolis, USA). All samples were tested in duplicate and values were corrected by running samples with culture media without cells to assess background NOx levels.

Piñero P., Juanola O., Caparrós E., Zapater P., Giménez P., González-Navajas J.M., Such J, & Francés R. (2017). Toll-like receptor polymorphisms compromise the inflammatory response against bacterial antigen translocation in cirrhosis. Scientific Reports, 7, 46425.

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Cytokine and Anti-Drug Antibody Levels

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The presence of anti-drug antibodies and serum levels of TNF-α, interferon-γ, and interleukin (IL)-12p40, IL-10, and IL-26 were determined by enzyme-linked immunosorbent assays (ELISAs) using Human Quantikine kits from R&D Systems (Minneapolis, MN, United States). ELISAs were also carried out to measure free infliximab and adalimumab levels and to detect anti-drug antibodies (Matriks Biotek, Ankara, Turkey), according to the manufacturer’s instructions. All samples were tested in triplicates and read in a Sunrise microplate reader (Tecan, Männedorf, Switzerland). The detection limit for each cytokine assay varied between 2 and 5 pg/ml and between 0.1 and 0.3 μg/ml in the case of free anti–TNF-α kits. Standard curves were generated for every plate, and the average z standard optical densities were subtracted from the rest of the standards and samples to obtain a corrected concentration for all parameters. The presence of anti-drug antibodies was evaluated by a cutoff value estimated by multiplying the optical density (OD) of the zero standard by 3, as indicated by the manufacturers. Samples were considered positive when the ratio sample OD/0 standard OD was higher than 3.

Orts B., Gutierrez A., Madero L., Sempere L., Frances R, & Zapater P. (2022). Clinical and Immunological Factors Associated with Recommended Trough Levels of Adalimumab and Infliximab in Patients with Crohn’s Disease. Frontiers in Pharmacology, 12, 795272.

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Quantification of Angiogenic Factors

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VEGF-A (reference DVE00), PDGF-BB (reference DBB00) and Angiopoietin-2 (reference DANG20) secreted in the ECFC and MSC culture medium or in patients’ plasma were quantified using the Human Quantikine kits (R&D Systems enzyme-linked immunosorbent assay) as previously described (22 (link)).

Smadja D.M., Levy M., Huang L., Rossi E., Blandinières A., Israel-Biet D., Gaussem P, & Bischoff J. (2015). Treprostinil indirectly regulates endothelial colony forming cell angiogenic properties by increasing VEGF-A produced by mesenchymal stem cells. Thrombosis and haemostasis, 114(4), 735-747.

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Cytokine Quantification Using ELISA Assay

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Levels of cytokines IL-1β, IL-6, TNFα, and IL-10 in cell culture supernatants were measured using Quantikine Human Kits (R&D Systems, Minneapolis, USA), according to the manufacturer's instructions. Protocol used was similar to that described by Jung et al. [22 (link)]. All reagents and working standards were prepared, and excess microplate strips were removed before adding 50 μL of RD1W (assay diluent) to each well. Next, 100 μL of standard control for the samples was added to each well, and the wells were covered with adhesive strips and incubated for 1.5 h at 25°C. Cells in each well were aspirated and washed twice (a total of three washes). Next, antiserum against each molecule to be analyzed was added to each well, and the wells were covered with new adhesive strips and incubated for 30 min at room temperature. The aspiration and washing steps were repeated. The conjugate (100 μL) was added to each well, and the plate was incubated for 30 min at room temperature. The aspiration and washing steps were repeated again. Next, 100 μL of substrate solution was added to each well, and the plate was incubated at room temperature for 20 min. The assay was terminated by adding 50 μL of stop solution to each well, and optical density was determined within 30 min by using a microplate reader set at 450 nm.

Barbisan F., Azzolin V.F., Teixeira C.F., Mastella M.H., Ribeiro E.E., do Prado-Lima P.A., Praia R.D., Medeiros Frescura Duarte M.M, & da Cruz I.B. (2017). Xanthine-Catechin Mixture Enhances Lithium-Induced Anti-Inflammatory Response in Activated Macrophages In Vitro. BioMed Research International, 2017, 4151594.

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Oxidative Stress and Cytokines in Rat Brains

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Once testing was completed, the rats were anesthetized with isoflurane and decapitated to extract the entire brain, which was dissected on ice. The amygdala, frontal cortex, hippocampus, and other brain structures were removed in that order, followed by the spleen and blood, all of which were frozen and stored at -80°C for analyses of the oxidative stress and cytokines.
Cytokines and 8'2-deoxyguanosine. The isolated brain structures were mechanically macerated with the aid of sterile slides and petri dishes containing Hanks' medium. The homogenate remained in an incubator for 2 h at 37ºC, with 5% CO 2 saturation. Then the material was centrifuged at 2000 rpm for 10 min, and the supernatant was removed and used as a sample for analysis of the cytokines and 8'2-deoxyguanosine. Tissue levels of the IL-1β, IL-6, TNF-alpha, IFN-γ, and IL-10 cytokines were measured using Quantikine Human Kits (R&D Systems, Minneapolis, USA) according to the manufacturer's instructions. DNA damage was determined by measuring 8'2-deoxyguanosine using an ELISA immunoassay kit obtained from Abcam (Cambridge, MA, USA) according to the manufacturer's instructions.

do Prado-Lima P.A.S., Onsten G.A., de Oliveira G.N., Brito G.C., Ghilardi I.M., de Souza E.V., Dos Santos P.G., Salamoni S.D., Machado D.C., Duarte M.M.F., Barbisan F., da Cruz I.B.M., Costa-Ferro Z.S.M, & daCosta J.C. (2019). The antidepressant effect of bone marrow mononuclear cell transplantation in chronic stress. Journal of psychopharmacology (Oxford, England), 33(5).

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