Total CD4+ T cells were isolated from PBMCs of HIV-infected individuals by negative selection using the CD4+ T Cell Isolation Kit (MACS, Miltenyi Biotec) on MACS LS columns. This kit contains the cocktail of monoclonal antibodies against CD8, CD14, CD15, CD16, CD19, CD36, CD56, CD123, TcRγ/δ, and CD235a. Total CD4+ T cells were then stained with APC anti-human CD32 antibody (clone FUN-2, SONY). CD32+CD4+ T cells were subsequently magnetically labeled with anti-APC microbeads (MACS, Miltenyi Biotec) and isolated by positive selection on MACS MS columns. Sorted CD32+CD4+ and CD32−CD4+ T cell fractions were either directly used for HIV DNA and RNA measurements or for the HIV reactivation assay (see below). The relative sizes of the sorted CD32+CD4+ and CD32−CD4+ T cell fractions were determined by cellular DNA quantitation in these fractions by qPCR using β-actin detection reagents (Thermo Fisher Scientific) and expressed as percentages of total CD4+ T cells.
Anti apc microbeads macs
Manufactured by Miltenyi Biotec
Anti-APC microbeads (MACS) are a laboratory tool used for the magnetic separation and isolation of cells expressing the APC (Allophycocyanin) antigen. These microbeads are designed to bind to the APC-labeled cells, allowing for their separation from the rest of the cell population using a MACS (Magnetic-Activated Cell Sorting) system.
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2 protocols using anti apc microbeads macs
1
Isolation and Characterization of CD32+ CD4+ T Cells
2
Isolation and Characterization of CD32+ CD4+ T Cells
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Total CD4+ T cells were isolated from PBMCs of HIV-infected individuals by negative selection using the CD4+ T Cell Isolation Kit (MACS, Miltenyi Biotec) on MACS LS columns. This kit contains the cocktail of monoclonal antibodies against CD8, CD14, CD15, CD16, CD19, CD36, CD56, CD123, TcRγ/δ, and CD235a. Total CD4+ T cells were then stained with APC anti-human CD32 antibody (clone FUN-2, SONY). CD32+CD4+ T cells were subsequently magnetically labeled with anti-APC microbeads (MACS, Miltenyi Biotec) and isolated by positive selection on MACS MS columns. Sorted CD32+CD4+ and CD32−CD4+ T cell fractions were either directly used for HIV DNA and RNA measurements or for the HIV reactivation assay (see below). The relative sizes of the sorted CD32+CD4+ and CD32−CD4+ T cell fractions were determined by cellular DNA quantitation in these fractions by qPCR using β-actin detection reagents (Thermo Fisher Scientific) and expressed as percentages of total CD4+ T cells.
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