Naive CD4 + T cells were infected with adenoviruses and differentiated in Th17 cell differentiation medium for 72 h. The levels of IL-17A in Th17 cells of mice were measured using an IL-17A ELISA kit (M17AF0, R&D Systems) according to the manufacturer’s instructions. The serum levels of IL-17A in human were measured according to the instructions provided in the ELISA kit (D1700, R&D Systems).
D1700
Manufactured by R&D Systems
Sourced in United States, Denmark
The D1700 is a compact and versatile laboratory instrument designed for centrifugation. It features a high-speed motor and a rotor with multiple tube positions for efficient sample processing.
Automatically generated - may contain errors
Lab products found in correlation
Dif50, by R&D Systems (2 mentions)
D6050, by R&D Systems (2 mentions)
Dta00c, by R&D Systems (1 mentions)
E el h0134c, by Elabscience (1 mentions)
Db100b, by R&D Systems (1 mentions)
D4050, by R&D Systems (1 mentions)
Dif50c, by R&D Systems (1 mentions)
Dta00d, by R&D Systems (1 mentions)
Dlb50, by R&D Systems (1 mentions)
Dmp100, by R&D Systems (1 mentions)
9 protocols using d1700
1
Measuring Th17 Cell-Derived IL-17A
2
Cytokine Quantification by ELISA
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The ex vivo culture medium was collected for subsequent enzyme-linked immunosorbent assay analysis according to the manufacturer’s instructions. In brief, 100 μL samples and standard were added to indicated wells and incubated for 2 hours at room temperature. Then, 100 μL biotin antibody was added and incubated for 1 hour at room temperature after incubation with streptavidin solution and TMB One-Step Substrate Reagent for 30 minutes, respectively. The reaction was terminated by Stop Solution and read at 450 nm immediately. To calculate the final concentration of indicated cytokine, ELISACalc software was used to generate standard curves. The enzyme-linked immunosorbent assay kits used in this study are as follows: DTA00C, DIF50, and D1700 (R&D Systems).
3
Quantifying Cytokines and MMP13 by ELISA
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The cytokines IL-17A and IL-10 (D1700 and D1000, respectively, purchased from R&D systems, Minneapolis, United States) and MMP13 (E-EL-H0134c, Elabscience Biotechnology, Wuhan, China) were determined using enzyme-linked immunosorbent assay (ELISA). The ELISA was performed according to the manufacturer’s instructions.
4
Quantification of Cytokine Levels
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The concentrations of IL-8, interferon (IFN)-γ, IL-4, IL-17A and TGF-β1 in cell culture supernatants were measured using Quantikine enzyme-linked immunosorbent assay (ELISA) kits for IL-8 (R&D Systems, D8000C), IFN-γ (R&D Systems, DIF50C), IL-4 (R&D Systems, D4050), IL-17A (R&D Systems, D1700) and TGF-β1 (R&D Systems, DB100B), respectively, according to the manufacturer’s instructions.
5
ELISA Kits for Cytokine Quantification
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Commercial ELISA kits (cat. nos. DTA00D, DLB50, D6050 and D1700; R&D Systems Inc.) were purchased to detect the serum levels of TNF-α, IL-1β, IL-6 and IL-17. All procedures, including sample and reagent preparation, assay procedure and calculation of the results were implemented according to the protocols of the kits.
6
Quantifying Cytokine Levels in Mouse LG and hCECs
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Protein levels of cytokines in mouse LG tissues and in culture supernatant of hCECs were quantified using commercially available ELISA kits: human IL-17 and IL-22 (D1700 and D2200, respectively; R&D Systems), human IL-1β, IL-6, IL-8, IL-23, TNF-α (437005, 430505, 431505, 435407, and 430205, respectively; BioLegend) and mouse IL-6, IL-17, IL-22 and IL23 (431305, 436205, 436305, and 433704, respectively; BioLegend).
7
Cytokine Secretion Measurement in Blood
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Supernatants from peripheral venous blood samples of patients and volunteers and the cell cultures were collected and stored at -80°C until the cytokine measurements. The secretions of IL-17A (R&D Systems, Inc., Cat# D1700) by coculture systems and IL-10 (R&D Systems, Inc., Cat# EL217B) by Tregs were measured in duplicate by ELISA following the manufacturer's recommendation. Results were expressed for each subject as cytokine concentration in pictogram per milliliter.
8
Quantifying Cytokine Levels in Mouse LG and hCECs
9
Quantification of Cytokines and MMP-1
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Culture supernatants were collected and kept at -20°C until use. Quantikine ELISAs were performed for the cytokines CXCL8, IL-6, IL-17A, IL-17B, IL-17C, IFN-γ, and for the metalloproteinase pro-MMP-1, as described by the manufacturer (D8000C, D6050, D1700, DY1248, DY1234, DIF50, DMP100, R&D Systems, Denmark). Briefly, human standard and samples were added to a 96-well plate and incubated at room temperature, washed and incubated with conjugate for 2 hours at room temperature. The plate was washed and a substrate was added. Optical density was read at 450 nm with a Tecan Infinite F200 reader (Labsystems Multiskan MS, USA). Sample concentrations were calculated from the standard sigmoid curve.
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