Anti mac 1 pe

BMMNCs were incubated for 45 minutes at 4°C with saturating concentrations of anti-mouse antibodies in ∼100 μL 3% FBS/0.09% NaN₃ in PBS with 0.25 μg anti-mouse CD16/CD32 (“Fc Block”). For the myeloid progenitor analysis, the following antibodies from BD Biosciences were used: FITC-conjugated anti-lineage markers (CD3, CD4, CD8, B220, Mac1, Gr1, Ter119), anti-CD16/32-PE, anti-CD34-PacificBlue, anti-Sca1-APC-Cy7, and anti-c-Kit-PerCP-Cy5.5. For the mature lineage analysis, the following antibodies from BD Biosciences were used: anti-CD3-FITC, anti-CD8-PacBlue, anti-B220-V500, anti-Mac1-PE, anti-Gr1-PECy7, anti-CD4-APC-Cy7, anti-CD45.2-PerCP-Cy5.5. Cells were analyzed on an LSR II 407 flow cytometer, and single color compensation controls were acquired using polystyrene microbeads (BD Biosciences). All post-acquisition analyses were performed with FlowJo 7.6.3 software (TreeStar, WA) with gating parameters determined by fluorescence minus-one controls.
The following gating definitions were used: MPs, myeloid progenitors; GMPs, granulocyte-monocyte progenitors; MEPs, megakaryocyte-erythroid progenitors. MPPs: Lin^–Sca1⁺c-Kit⁺; MPs: Lin^–Sca1^–c-Kit⁺. CMPs: Lin^–Sca1^–c-Kit⁺CD34⁺FcγRII/II^–/lo. GMPs: Lin^–Sca1^–c-Kit⁺CD34⁺FcγRII/ II⁺. MEPs: Lin^–Sca1^–c-Kit⁺CD34^–FcγRII/II^–. HSPCs: CD150⁺Lin^–CD48^–CD41^–Sca1⁺c-Kit⁺. Myeloid cells: CD3^–B220^–; Monos: CD3^–B220^–Gr1^–Mac1⁺.