The CXCL12 baseline serum levels of 108 patients suffering from RA were assessed with a quantitative sandwich ELISA kit (R&D Systems, Minneapolis, MN, USA; sensitivity 47 pg/ml). Analysts were blinded with regard to ADA status of the patients.
Quantitative sandwich elisa kit
Manufactured by R&D Systems
Sourced in United States
The Quantitative sandwich ELISA kit is a laboratory equipment designed for the quantitative detection and measurement of target analytes in biological samples. It utilizes the sandwich ELISA principle, where the target analyte is captured between two specific antibodies. The kit provides the necessary reagents and consumables for performing the assay, enabling accurate and reproducible quantification of the analyte of interest.
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Lab products found in correlation
Ensight multimode plate reader, by PerkinElmer (1 mentions)
Cobas integra 400 plus analyzer, by Roche (1 mentions)
Path scan elisa kit, by Cell Signaling Technology (1 mentions)
Quantikine hs elisa human il 6 immunoassay, by R&D Systems (1 mentions)
Quantikine hs elisa human tnf α immunoassay, by R&D Systems (1 mentions)
High sensitivity elisa kit, by R&D Systems (1 mentions)
14 protocols using quantitative sandwich elisa kit
1
CXCL12 Levels in Rheumatoid Arthritis
2
Quantification of Inflammatory Cytokines
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TNF-α levels were measured in human cultured lymphocytes after the treatment described above by using highly sensitive TNF-α ELISA (R & D Systems, Minneapolis, MN, USA) according to the manufacturer’s instructions. Pro-inflammatory cytokine (IL-1β and IL-6) levels were determined with a quantitative sandwich ELISA kit (R & D Systems, Minneapolis, MN, USA) according to the manufacturer’s instructions [15 (link)]. The reaction was developed with streptavidin-horseradish peroxidase and optical density was read at 450 nm wavelength with the Perkin Elmer EnSight Multimode Plate Reader.
3
Isolation and Quantification of Human Monocyte MMPs
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To obtain human monocytes, peripheral blood mononuclear cells were seeded in petri dishes at the density of 106/mL. The cells were allowed to adhere to plastic tissues and non-adherent cells (lymphocytes) were removed. In cultured monocytes, MMP levels were measured after the treatment described above by using the corresponding quantitative sandwich ELISA kit (R & D Systems) according to the manufacturer’s instructions [15 (link)]. Briefly, the assay systems measure natural and recombinant human active and pro-MMPs (total MMPs).
4
Plasma CysC and Urinary Biomarkers Measurement
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Plasma CysC was measured by a commercially available quantitative sandwich ELISA kit (R&D Systems, MN, USA) for mice according to the manufacturer’s instructions. In brief, plasma samples were diluted 200-fold and incubated in a 96 well microplate pre-coated with CysC specific antibody and CysC concentration was determined from the color intensity of oxidized Tetramethylbenzidine (TMB) measured at 450 nm. Urinary total protein, creatinine and plasma blood urea nitrogen were measured by COBAS INTEGRA 400 plus analyzer (Roche Diagnostics, Rotkreuz, Switzerland). To measure urinary Na+, samples were diluted 1500-fold with 1.0% v/v trace metal free nitric acid and analyzed by using a Varian VISTA AX CCD Simultaneous Inductively Coupled Plasma Atomic Emission Spectroscopy (Varian, CA, USA). Standards Na+ for Inductively Coupled Plasma Mass Spectrometry (Spex CertiPrep, NJ, USA) was used to make standards ranging from 0.05 ppm to 5.0 ppm. Certified urine controls (Seronorm, Stasjonsveien, Norway) and sample pool controls were used to check the accuracy of the assay.
5
Quantification of VEGF and STAT3 in Cells
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Total VEGF in the conditioned medium of cultured cells was measured using quantitative sandwich ELISA kit (R&D Systems) according to the manufacturer's instructions. Path scan ELISA kit (Cell Signaling Technologies) using STAT3 and p-STAT3 (Tyr 705) coated antibodies was according to the manufacturer's instructions. The data is represented as absorbance units at 450 nm from at least three independent experiments. Data are represented as mean ± standard deviation of the mean (SD).
6
Quantification of IGF1, Leptin, and PTX3
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IGF1 and leptin levels were quantified in the conditioned media from 1 × 105 MSCs using a quantitative sandwich ELISA kit (Fine Biotech, China). The PTX3 level was quantified in the same way from the obtained media using a quantitative sandwich ELISA kit (R&D Systems).
7
Quantitative Assay of Leptin and VEGF
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Leptin and VEGF assays were conducted centrally at Medpace, Inc., Cincinnati, Ohio, based on samples collected at three times: (1) baseline; (2) at follow‐up years 8, 10, or 12; and (3) at follow‐up years 14, 16, or 18. Serum leptin concentrations were measured by a monoclonal antibody‐based quantitative sandwich enzyme‐linked immunosorbent assay (ELISA) kit (R&D Systems, Inc., Minneapolis, MN) fully validated in the laboratory. The assay low level of quantification was 1.56 ng/mL, and the reference range was 1.0 to 50.0 ng/mL. Three quality control samples with low, medium, and high levels of leptin were analyzed in each plate, and the interassay coefficients of variation were typically < 6.0% for all three control samples.
Serum VEGF concentrations were measured by a monoclonal antibody‐based quantitative sandwich ELISA kit (R&D Systems) that was fully validated in the laboratory. The assay low limit of quantification was 50.0 pg/mL, and the reference range was 62 to 707 pg/mL. Three quality control samples with low, medium, and high levels of VEGF were analyzed in each plate and the interassay coefficients of variation were typically 12.4%, 9.5%, and 10.6%, respectively.
Serum VEGF concentrations were measured by a monoclonal antibody‐based quantitative sandwich ELISA kit (R&D Systems) that was fully validated in the laboratory. The assay low limit of quantification was 50.0 pg/mL, and the reference range was 62 to 707 pg/mL. Three quality control samples with low, medium, and high levels of VEGF were analyzed in each plate and the interassay coefficients of variation were typically 12.4%, 9.5%, and 10.6%, respectively.
8
MCAO-Induced Inflammatory Markers in Serum
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Blood samples were collected at 24 h after MCAO. Whole blood was centrifuged (13000 ×g for 15 min) and supernatants were collected to determine the level of TNF-α, IL-1β, and IL-6 in serum by available quantitative sandwich ELISA kits (R&D, USA). All use of ELISA kits was in strict accordance with the manufacturer's protocols. The concentrations of the samples were calculated according to the standard curve. The serum TNF-α, IL-1β, and IL-6 levels were all expressed as ng/L.
9
VEGF and MMP-2/9 Serum Quantification
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Quantification of serum levels of VEGF and MMP-2/9 were assessed in triplicate by commercially available quantitative sandwich ELISA kits according to instructions of the manufacturers (R&D Systems, Minneapolis, MN). The results were expressed as picograms of VEGF per milliliter (pg/ml) or nanogram of MMP-2/9 per milliliter (ng/ml), based on the standards provided with the available kits.
10
Cytokine response to CFCS treatment
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Cells were cultured in the recommended media until they reached 80% cell density. After that, cells were treated with CFCS at various concentrations (2.5, 5, and 10 mg/mL) and incubated for 24 h. The levels of interleukins IFN-γ and IL-6 were estimated using commercially available quantitative sandwich ELISA kits (R&D Systems, Inc, Minneapolis, MN, USA), according to the manufacturer’s protocol. A 450 nm wavelength was used to read the plates. The results were expressed as milligrams of protein per plate.
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