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Multiplex Cytokine/Chemokine Assay

JJ Junseok Jeon
KL Kyungho Lee
KY Kyeong Eun Yang
JL Jung Eun Lee
GK Ghee Young Kwon
WH Wooseong Huh
DK Dae Joong Kim
YK Yoon-Goo Kim
HJ Hye Ryoun Jang
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Multiplex cytokine and chemokine analysis in whole kidney protein extracts was conducted using a Milliplex MAP Mouse Cytokine/Chemokine Kit (Luminex, Austin, TX) following the manufacturer's instructions. Anti-cytokine monoclonal antibodies linked to microspheres incorporating distinct properties of two fluorescent dyes were used in this multiplexed particle-based flow cytometric assay. Our assay was designed to quantify interleukin (IL)-2; IL-4; IL-6; IL-10; interferon (IFN)-γ; monocyte chemoattractant protein (MCP)-1; regulated on activation, normal T cell expressed and secreted (RANTES/CCL5); tumor necrosis factor (TNF)-α; and vascular endothelial growth factor (VEGF). The value of each cytokine or chemokine was normalized by dividing the raw concentration (pg/ml) by the kidney protein concentration (mg/ml, measured by using a Pierce BCA protein assay kit, Thermo Fisher Scientific, Waltham, MA).

Copyright and License information:  ©2021 Jeon, Lee, Yang, Lee, Kwon, Huh, Kim, Kim and Jang.
This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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