Anti rat cd45 fitc

SVF cells from RPAT pads from CTR and MSG animals were isolated, and at least 2 × 10⁵ cells (in 100 µL Phosphate-buffered saline (PBS)/0.5% Bovine Serum Albumin (BSA)) were incubated with fluorescent antibodies or respective isotype controls (1/50 diluted, for 1 h at 4 °C). After washing steps, flow cytometry was analyzed using a FACSCalibur flow cytometer (Becton Dickinson Biosciences, San Jose, CA, USA). A combination of cell surface markers was used to identify APCs as: CD34⁺/CD45⁻/CD31⁻ [31 (link)]. The conjugated monoclonal antibodies used were: anti-rat CD34:PE (PE: phycoerythrin), anti-rat CD45:FITC (FITC: fluorescein isothiocyanate) and anti-rat CD31:FITC (1 µg/1 × 10⁶ cells, Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA). Samples were analyzed using CellQuest Pro (Becton-Dickinson, San Jose, CA, USA) and FlowJo software (TreeStar, San Carlo, CA, USA).

Freshly isolated or cultured SVF cells from RPAT pads (at least 2 × 10⁵ cells in 100 µL PBS/0.5% BSA) were incubated with fluorescent antibodies or respective isotype controls for 1 h at 4 °C. After washing steps, flow cytometry was performed using a FACSCalibur flow cytometer (Becton Dickinson Biosciences, San Jose, CA, USA). A combination of cell surface markers were used to identify APCs as: CD34⁺/CD45⁻/CD31⁻ for freshly isolated SVF cells and CD34⁺/CD31⁻ for cultured SVF cells [22 (link)]. Conjugated monoclonal antibodies used were: anti-rat CD34:PE, anti-rat CD45:FITC, and anti-rat CD31:FITC (1 µg/1 × 10⁶ cells, Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA). Samples were analyzed using CellQuest Pro (Becton-Dickinson, San Jose, CA, USA) and FlowJo softwares (TreeStar, San Carlo, CA, USA).

SVF cells from RPAT pads of C and F animals were isolated and at least 2 × 10⁵ cells (in 100 µL PBS/0.5% BSA) incubated with fluorescent antibodies or respective isotype controls for 1 h at 4 °C. After washing steps, flow cytometry was analyzed using a FACS Calibur flow cytometer (Becton Dickinson Biosciences). A combination of surface cell markers were used to identify adipocyte precursor cells (APCs) as: CD34⁺/CD45⁻/CD31⁻ [27 (link)]. Conjugated monoclonal antibodies used were: anti-rat CD34:PE (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA), anti-rat CD45:FITC (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA) and anti-rat CD31:FITC (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA). Samples were analyzed using CellQuest Pro (Becton-Dickinson, San Jose, CA, USA) and FlowJo softwares (TreeStar, San Carlo, CA, USA) [27 (link)].