Cells were sorted using FACS or magnetic-activated cell sorting (MACS). For FACS sorted cells, red blood cell lysed splenocytes were stained with the following antibody cocktail: APF-Cy7 labeled anti-B220 (Biolegend; San Diego, CA), APC labeled anti-CD3 (Life Technologies Corp; Carlsbad, CA), and BV421 labeled anti-F4/80 (Biolegend; San Diego, CA). For MACS sorted cells, red blood cell lysed splenocytes were labeled using CD4 and CD8 antibody conjugated microbeads (Miltenyi Biotec; Bergisch Gladbach, Germany) and positively sorted as detailed by the manufacturer. NK cells were negatively sorted per manufacturer’s instructions using an NK cell isolation kit (Miltenyi Biotec; Bergisch Gladbach, Germany). RNA from sorted cells was isolated and gene expression analyzed as described below.
Cd4 and cd8 antibody conjugated microbeads
Manufactured by Miltenyi Biotec
Sourced in Germany
CD4 and CD8 antibody conjugated microbeads are laboratory reagents used for the isolation and enrichment of CD4+ and CD8+ T cells from biological samples. These magnetic beads are coated with monoclonal antibodies specific to the CD4 and CD8 cell surface markers, allowing for the selective separation of these cell populations.
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2 protocols using cd4 and cd8 antibody conjugated microbeads
1
Cell Sorting and Gene Expression Analysis
2
Cell Sorting and Gene Expression Analysis
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Cells were sorted using FACS or magnetic-activated cell sorting (MACS). For FACS sorted cells, red blood cell lysed splenocytes were stained with the following antibody cocktail: APF-Cy7 labeled anti-B220 (Biolegend; San Diego, CA), APC labeled anti-CD3 (Life Technologies Corp; Carlsbad, CA), and BV421 labeled anti-F4/80 (Biolegend; San Diego, CA). For MACS sorted cells, red blood cell lysed splenocytes were labeled using CD4 and CD8 antibody conjugated microbeads (Miltenyi Biotec; Bergisch Gladbach, Germany) and positively sorted as detailed by the manufacturer. NK cells were negatively sorted per manufacturer’s instructions using an NK cell isolation kit (Miltenyi Biotec; Bergisch Gladbach, Germany). RNA from sorted cells was isolated and gene expression analyzed as described below.
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