Figure 1

Paw Cytokine Extraction Method Flow Sheet. As the illustration demonstrates, paws are selected for processing, snap frozen and stored at -80°C until ready to be processed. When ready, samples are placed on dry ice and homogenized frozen using prepared tissue extraction buffer. An aliquot is used for Luminex^® cytokine analysis, phosphorylated STAT (pSTAT) pharmacodynamic (PD) Luminex^® analysis and another aliquot used for pharmacokinetic (PK) analysis. After all three analyses are finished; the PD data is compared to the functional cytokine readouts. PK data should always be compared against the functional PD data to see if pharmacological saturation of the target has been achieved. Thus, multiple cytokines and intracellular signaling molecules can be monitored using multiplex bead technology to better predict local disease outcomes and help design more targeted therapies.