The experimental treatments and groupings were the same as those used in 2.7, the HFLS in the logarithmic growth phase were seeded in 96-well culture plate at 5 × 104 cells per well. After 72 h of TGF-β1 induction, the supernatants were collected and centrifuged at 10,000×g and 4 °C for 2 min, and then were used to detect the inflammatory factors TNF-α, IL-1β and IL-6 following the instructions of ELISA kits (ES24RB, EH2IL6 and BMS224-2, Thermofisher) respectively.
Eh2il6
Manufactured by Thermo Fisher Scientific
Sourced in United States
The EH2IL6 is a laboratory equipment product from Thermo Fisher Scientific. It is designed to perform specific functions in a research or diagnostic setting. The core function of this equipment is to provide precise and reliable results for users. However, without further details on the specific capabilities and intended use of this product, a more detailed description cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Khc3011, by Thermo Fisher Scientific (5 mentions)
Bms224 2, by Thermo Fisher Scientific (4 mentions)
Es24rb, by Thermo Fisher Scientific (2 mentions)
Kac1751, by Thermo Fisher Scientific (2 mentions)
E el h0088c, by Elabscience (1 mentions)
Bms215 2, by Thermo Fisher Scientific (1 mentions)
Bms223 4, by Thermo Fisher Scientific (1 mentions)
Alamarblue cell viability assay, by Bio-Rad (1 mentions)
Bms224hs, by Thermo Fisher Scientific (1 mentions)
Kac1591, by Thermo Fisher Scientific (1 mentions)
Khc0081, by Thermo Fisher Scientific (1 mentions)
14 protocols using eh2il6
1
Assaying Inflammatory Factors in TGF-β1-Treated Cells
2
Cytokine Quantification in Tissue and Culture
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The levels of TNF-α (KHC3011, Thermo, USA), IL-6 (EH2IL6, Thermo, USA), and IL-1 β (E-EL-H0088c, Elabscience, USA) in tissue lysates and cell culture medium were detected by using commercial ELISA assay kit following manufacturer's protocols.
3
Quantifying Inflammatory Cytokines by ELISA
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Enzyme-linked immunosorbent assays (ELISA) were utilized for the quantification of inflammatory antigens in the cell supernatant. The assessed markers included human Interleukin (IL)-1β, IL-4, IL-6, IL-10 and Tumor necrosis factor α (TNF-α). All ELISA kits were obtained from Thermo Fisher Scientific, with the following kit numbers: BMS224-2 (IL-1β), BMS225-2 (IL-4), EH2IL6 (IL-6), BMS215-2 (IL-10) and BMS223-4 (TNF-α). All reagents were maintained at room temperature before use. Cytokine markers were detected at a wavelength of 450 nm, as per manufacturer’s protocols. A standard curve was used for each assay to determine the cytokine concentration levels of the samples, as per manufacturer’s protocols and expressed in pg/mL.
4
Quantification of Inflammatory Mediators
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According to the manufacturer’s instructions, the concentrations of interleukin (IL)-1β (BMS224–2, Thermofisher), IL-6 (EH2IL6, Thermofisher), TNF-α (KHC3011, Thermofisher), and CGRP (ABIN1095216, Antibodies-online) in the cell culture supernatants were analyzed using an ELISA kit. An automated microplate reader (SpectraMax® M5) was used for the measurement of the optical density (OD) at 450 nm. The concentrations of each sample were detected based on optical density and the concentration of the standard.
5
Quantifying IL6 Secretion in T47D Cells
6
Phosphorylated Src and Cytokine ELISA
7
Measuring Serum IL-6 Levels by ELISA
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Serum IL-6 concentration was measured by a specific
human IL-6 ELISA kit (EH2IL6, Thermo Fisher Scientific, USA). In this method, 50 µl of serum is loaded into the
wells containing IL-6 antibody. Then a washing step was
performed to wash the other analytes. In the next step, a
substrate was added, resulting in a blue color development
proportional to the amount of IL-6 in the serum. Finally,
the reaction was stopped by adding the stop solution, and
the amount of yellow color was assessed at a wavelength
of 450 nm by an ELISA reader (DRG instrument, Cat. No.
ELM-2000, Germany).
human IL-6 ELISA kit (EH2IL6, Thermo Fisher Scientific, USA). In this method, 50 µl of serum is loaded into the
wells containing IL-6 antibody. Then a washing step was
performed to wash the other analytes. In the next step, a
substrate was added, resulting in a blue color development
proportional to the amount of IL-6 in the serum. Finally,
the reaction was stopped by adding the stop solution, and
the amount of yellow color was assessed at a wavelength
of 450 nm by an ELISA reader (DRG instrument, Cat. No.
ELM-2000, Germany).
8
IL-6 ELISA Protocol: Media Removal
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Media at the indicated time points and conditions were removed prior to harvesting cells, diluted in the assay buffer, and subjected to ELISA (Thermo Fisher, EH2IL6) as recommended by the manufacturer’s using recombinant human IL6 supplied for generating a standard curve.
9
Evaluating Anti-Inflammatory Effects of CPP-AIF
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Macrophage inflammatory assay were carried out to evaluate whether CPPAIF displayed anti-inflammation potential [22 (link),23 (link),24 (link)]. In this experiment, macrophage (Raw264.7) cells were seeded in 96-well plates (5 × 105 cells/mL) and allowed to attach overnight. After attachment, cells were incubated with various concentrations of CPPAIF for 1 h and followed by stimulation with 1 μg/mL of LPS (lipopolysaccharide). No LPS-added cells were considered as control groups. After incubation, the amount of TNF-α and IL-6 in the medium were analyzed by enzyme linked immunosorbent assay (# KHC3011 and #EH2IL6, Thermo Fisher, Waltham, MA, USA). Cell viability was measured by AlamarBlue cell viability assay (BUF012B, Bio-Rad, Hercules, CA, USA).
10
Assaying Inflammatory Factors in TGF-β1-Treated Cells
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The experimental treatments and groupings were the same as those used in 2.7, the HFLS in the logarithmic growth phase were seeded in 96-well culture plate at 5 × 104 cells per well. After 72 h of TGF-β1 induction, the supernatants were collected and centrifuged at 10,000×g and 4 °C for 2 min, and then were used to detect the inflammatory factors TNF-α, IL-1β and IL-6 following the instructions of ELISA kits (ES24RB, EH2IL6 and BMS224-2, Thermofisher) respectively.
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