Cell culture supernatants in the four groups were collected for cytokine evaluation. The level of IL-6, IL-10, TNF-α, TGF-β1 were measured by enzyme immunoassay kit (Bosterbio, Wuhan, People’s Republic of China). The content of cytokines was calculated through a standard curve obtained by measuring the standards.
Enzyme immunoassay kit
Manufactured by Boster Bio
Sourced in China
The enzyme immunoassay kit is a laboratory tool used for the detection and quantification of specific proteins or other analytes in a sample. It utilizes the principle of antigen-antibody interactions and enzyme-catalyzed reactions to provide a sensitive and accurate measurement.
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Lab products found in correlation
4 protocols using enzyme immunoassay kit
1
Cytokine Evaluation in Cell Culture
2
Cytokine Quantification in Macrophages
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According to the instructions, the levels of TNF-α, IL-1β, and IL-6 were detected using an Enzyme Immunoassay Kit (Boster, Inc., Wuhan, China). Briefly, when the macrophages reached 70% fusion, the cell supernatant was collected, the antigen was added and kept at 4°C overnight, and then enzyme-labeled secondary antibody (Abcam Inc., Cambridge, MA, USA) was added and incubated at room temperature for 1 h. Finally, the optical density of each sample at 450 nm was measured, and the levels of TNF-α, IL-1β, and IL-6 were calculated according to the regression equation of the standard curve.
3
VEGF and PGE2 Quantification in Peritoneal Fluid
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Peritoneal fluid was collected by rinsing the abdominal cavity with 10 mL of PBS and immediately centrifuged at 1500 rpm during 10 min. Supernatants were stored at -70°C until assayed for VEGF and PGE2 by use of an enzyme immunoassay kit (Boster Biological Technology, Pleasanton, CA and Cayman Chemical, Ann Arbor, MI), according to manufacturer's instructions. The concentrations were calculated from standard curves and all samples were assessed in triplicate. The VEGF and PGE2 measurement were performed by an automatic plate reader (Spectra Max; Molecular Devices, Sunnyvale, Calif) controlled by SoftMax software (Molecular Devices).
4
Transfection of HUVECs with miRNA Mimics and Inhibitors
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HUVECs were transfected with control miRNA, miR-181a-5p, or miR-181a-3p mimics or their inhibitors. Twenty-four hours later, cells were treated with 10 ng/ml TNF-α for 24 h. The cultured medium were then collected and ICAM-1, VCAM-1, and E-selectin levels were measured by ELISA analysis using Enzyme Immunoassay Kit (Boster, Inc. Wuhan, China) according to the instructions of the supplier.
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