The HSC preparation was a minor modification from previously described method.33 In brief, the whole bone marrow cells were collected from tibias in treated mice. Bone marrow cells were stained with antibodies for the identification of HSCs (c‐Kit+ Sca1+ Lin–), and the following antibodies were used: c‐Kit‐PE (#12‐1171‐82), Sca‐1‐FITC (#11‐5981‐82), and anti‐Lineage Antibody Cocktail comprises a mixture of PE‐Cy5‐conjugated antibodies, including anti‐B220, anti‐CD4, anti‐CD8, anti‐Gr‐1, anti‐Mac‐1, and anti‐TER119 (from eBioscience). For HSC sorting, debris, dead, and clumped cells were removed to obtain single and viable cells; then, the c‐Kit+ Sca1+ Lin– cell population was isolated by HSC sorting; FACS analysis was performed on BD FACSMelody™ Cell Sorter.34
Anti mac 1
Manufactured by Thermo Fisher Scientific
Anti‐Mac‐1 is a laboratory reagent used in flow cytometry applications. It is a monoclonal antibody that binds to the Mac‐1 (CD11b/CD18) antigen expressed on the surface of myeloid cells, including monocytes, macrophages, and granulocytes. The core function of Anti‐Mac‐1 is to facilitate the identification and analysis of these cell populations in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti gr 1, by Thermo Fisher Scientific (2 mentions)
Anti cd8, by Thermo Fisher Scientific (1 mentions)
Anti b220, by Thermo Fisher Scientific (1 mentions)
Facsmelody cell sorter, by BD (1 mentions)
Anti cd4, by Thermo Fisher Scientific (1 mentions)
Anti ter119, by Thermo Fisher Scientific (1 mentions)
Anti nk1, by Thermo Fisher Scientific (1 mentions)
Anti foxp3, by Thermo Fisher Scientific (1 mentions)
Anti c kit, by Thermo Fisher Scientific (1 mentions)
Lsr 2, by BD (1 mentions)
3 protocols using anti mac 1
1
Hematopoietic Stem Cell Isolation
2
Kinetic Analysis of Immune Cell Populations
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10 week old female B6.2 mice were treated daily for 0 to 10 days with 3AC as described above. On each day (0 to 10 inclusively), 5 mice were sacrificed, splenocytes were harvested, RBCs were lysed using 1 × RBC Lysis buffer (eBioscience), counted and then stained with anti-NK1.1, anti-Mac1 and anti-Gr1, or with anti-CD3ε and anti-CD4 then fixed and stained with anti-FoxP3 as per manufacturer's recommendation (eBioscience). Cells were analyzed by flow cytometry as described below. Frequency and absolute numbers of live NK cells and MDSCs were compared to those observed in the splenocytes from the 5 uninjected mice harvested on day 0. Live CD4+FoxP3+ cells were expressed as frequency of CD3ε+ T-cells and compared to those on day 0.
3
Immunophenotyping of Immune Cells
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For FACS analysis, cells were stained with antibodies in staining buffer (1× PBS, 2% FBS) and incubated at 4°C for 30 minutes. The samples were washed once with staining buffer before subjected to FACS analysis with the use of a BD LSRII. The antibodies used in this study include anti–Mac-1(eBioscience), anti–Gr-1(eBioscience), anti–c-KIT (eBioscience).
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