IRAK4 degradation in blood by FLOW
FLOW cytometry was performed at CellCarta. Whole blood samples were collected in Na-Hep tubes at the clinical site and shipped on the day of collection at 4 °C. Upon receipt, samples were lysed/fixed and stored at −80 °C until processing. Lysed/fixed samples were divided into plates for unblocked and blocked conditions, stained with a pre-permeabilization antibody cocktail (CD14 1:25, CD56 1:50, CD19 1:100, CD3 1:200, CD4 1:200, CD8 1:200, CD45 1:200, CD15 1:200) for 30 min at room temperature, washed and permeabilized with 60% methanol at 4 °C for 10 min, followed by incubation with IRAK4 unconjugated antibody at 12.5ug ml