Rat igg2a kappa control pe

Grafts from the fresh SVF gel group, 1-month-frozen SVF gel group, fresh fat group, and 1-month-frozen fat group were harvested at 1 month after transplantation. Samples were digested, and the SVF was assessed by flow cytometry. Briefly, 0.5 cc of graft tissues was minced with scissors and incubated in 0.075% collagenase buffer in a shaking water bath (37 °C, 30 min) for digestion. After neutralization and filtration, SVF cells were washed and stained with the following antibodies and corresponding isotype controls: anti-CD11b-PE (Miltenyi Biotec, Bergisch Gladbach, Germany), anti-F4/80-APC (eBioscience, Inc., CA, USA), rat IgG2a Control APC (BD Biosciences), and rat IgG2a Kappa Control PE (BD Biosciences). Samples were analyzed by flow cytometry (MACSQuant Analyzer 10; Miltenyi Biotec). Gating for each signal was set to eliminate 99.9% of the cells in the corresponding isotype control. Experiments were conducted in triplicate using 0.5-cc grafts collected from three mice in the same groups.

For flow cytometry analysis, suspended ECFCs and ASCs were filtered, washed, and stained with the following antibodies and corresponding isotype controls: anti-CD45-Cy5 (Miltenyi Biotec, Bergisch Gladbach, Germany), anti-CD133-PE (eBioscience, Inc., CA, USA), anti-CD31-FITC (BD Biosciences), anti-CD105-FITC (BD Biosciences), rat IgG2a Kappa cy5 (BD Biosciences), and rat IgG2a Kappa Control PE (BD Biosciences). Samples and controls were analyzed with a MACSQuant Analyzer 10 (Miltenyi Biotec). Each channel was gated respective to the control samples.