Anti-Myeloperoxidase antibody (ab9535)

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Publication protocol

Harvested samples of eWAT, iWAT and BAT were fixed in formalin and embedded in paraffin. Sections were stained using hematoxylin and eosin (H&E) and automated immunohistochemistry was performed (Ventana Medical Systems, Tucson, AZ) using anti-UCP1 and anti-myeloperoxidase (MPO) antibodies (Abcam, Cambridge, MA). Whole slide digital images were obtained from the resultant slides using an Aperio AT2 digital slide scanner (Leica Microsystems, Buffalo Grove, IL). The high resolution digital images were analyzed using Image Scope™ software (Aperio version 12) using the positive pixel count algorithm as described by Gannon et al. (21). In brief, the algorithm generates three classes of output values (weak positive, positive, and strong positive) based on pixel intensity which represents staining intensity. Only positive and strong positive values were included in the analysis. Three similar-sized areas were randomly selected on each digital slide and the average value of those three areas was used as the final value for comparison.

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