Rat anti ckit apc cy7

PB of mice was collected retro-orbitally, and hematopoietic parameters were measured by complete blood counts. Central bone marrow was flushed from femurs, and cellularity was quantified with 3% acetic acid in methylene blue (STEMCELL Technologies, 07060). For flow cytometric analysis of the lineage cell compartment of mice, flushed bone marrow cells were stained for myeloid (rat anti–Gr-1 APC-Cy7, BD Biosciences, 557661; rat anti–Mac-1 APC, eBioscience, 17-0112-83) or lymphoid lineages (rat anti-B220 FITC, eBioscience, 11-0452-82; hamster anti-CD3 PE-Cy7, eBioscience, 25-0031-82). The HSPC compartment was analyzed by staining for Lineage (biotin–Ter-119, –Mac-1, –Gr-1, -CD4, -CD8α, -CD5, -CD19, and -B220, eBioscience, 13-5921-85, 13-0051-85, 13-5931-86, 13-0112-86, 13-0452-86, 13-0041-86, 13-0081-86, 13-0193-86) followed by staining with streptavidin–PE-Texas Red (Invitrogen, SA1017), rat anti-cKit APC-Cy7 (eBioscience, 47-1171-82), rat anti-Sca1 PerCP-Cy5.5 (eBioscience, 45-5981-82), hamster anti-CD48 APC (eBioscience, 17-0481-82), and rat anti-CD150 PE-Cy7 (BioLegend, 115914). All flow cytometry analysis was performed on a BD LSR Fortessa flow cytometer with FlowJo v10.7.1 for Mac.

Peripheral blood of mice was collected retro-orbitally. To analyze the peripheral blood of mice, red blood cells were first lysed and then stained for myeloid (rat anti-Gr-1 APC-Cy7, BD, 557661, rat anti-Mac-1 APC, eBioscience, 17-0112-83) or lymphoid lineages (rat anti-B220 FITC, eBioscience, 11-0452-82, hamster anti-CD3 PE-Cy7, eBioscience, 25-0031-82).
Bone marrow was flushed from femurs and cellularity was quantified with 3% acetic acid in methylene blue (STEMCELL). The lineage cell compartment of the bone marrow was analyzed by staining for myeloid and lymphoid lineages as in the peripheral blood. The HSPC compartment was analyzed by staining for Lineage (biotin-Ter-119, -Mac-1, -Gr-1, -CD4, -CD8α, -CD5, -CD19 and -B220 (eBioscience, 13-5921-85, 13-0051-85, 13-5931-86, 13-0112-86, 13-0452-86, 13-0041-86, 13-0081-86, 13-0193-86) followed by staining with streptavidin-PE-TexasRed (Invitrogen, SA1017), rat anti-cKit APC-Cy7 (eBioscience, 47-1171-82), rat anti-Sca1 PerCP-Cy5.5 (eBioscience, 45-5981-82), hamster anti-CD48 APC (eBioscience, 17-0481-82) and rat anti-CD150 PE-Cy7 (Biolegend, 115914). All flow cytometry analysis was performed on a BD LSR Fortessa flow cytometer and was analyzed on FlowJo v10.5.3 for MAC.