After collagenase digestion and purification, the floating layer contains adipocytes, but also macrophages in the obese condition. Floating layer cells and stromal vascular cells (SVC) were collected and mRNA was extracted and reverse transcribed. qRT-PCR was performed using indicated primer pairs and relative expression levels were determined after normalizing to GAPDH. (a) Adiponectin, the marker for intact adipocytes, is detected in the floating layer, but not in the SVC layer indicating that the floating layer contains the adipocytes. (b) The macrophage markers CD68 and F4/80 indicate a significant increase in macrophage presence in the SVC layer and the floating layer of obese animals. Macrophages were also purified from SVC using magnetic bead separation linked to anti-F4/80 and mRNA for F4/80 quantitated.