Analysis of cytokine levels in lung tissue

LH Laura Hesse
AP Arjen H. Petersen
JE Joanne N. G. Oude Elberink
AO Antoon J. M. van Oosterhout
MN Martijn C. Nawijn
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The right superior lung lobe was used for measurement of total protein content and a cytokine profile. First, lungs were weighed, homogenized and dissolved in Luminex buffer (weight to volume ratio 1:5) and the total protein content was measured using a BCA protein assay according to manufacturer’s protocol (Thermo Scientific, USA). Concentrations of IL4, IL5, IL-10, IL13, IL-17, IL33, IFNγ, Eotaxin (CCL11), TARC (CCL17), and MIP3α (CCL20) were measured using a multiplex Mouse Cytokine/Chemokine Magnetic Bead Panels (MILLIPLEX Map Kit; Merck Millipore, Germany) according to manufacturer’s protocol. Plates were analyzed using a MAGPX1023 4002 with Luminex xMAP technology.

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