For flow cytometry analysis, in vitro cultured erythroid cells were washed in FACS buffer (1% FBS in PBS) before antibody staining. The following antibodies at indicated dilutions were used in this study: 1:50 APC-conjugated CD235a (Glycophorin A, clone HIR2, 50–153-69, eBioscience), 1:50 FITC-conjugated CD71 (OKT9, 14–0719-82, eBioscience), 1:50 APC-conjugated CD49d (9F10, Biolegend, 304308) and 1:200 FITC-conjugated BAND3 (An et al., 2014 (link); Hu et al., 2013 (link)). For live/ dead cell discrimination Sytox Blue was used according to the manufacturer’s instructions (Thermo Fisher, S34857). For apoptosis analysis, BV421-Annexin V (BD Bioscience, 563973) was used in combination with Sytox Orange according to the manufacturer’s instructions (Thermo Fisher, S34861). FACS analysis was conducted on a BD Bioscience Fortessa flow cytometer at the Whitehead Institute Flow Cytometry core. Data were analyzed using FlowJo software v10.4.2.
Apc conjugated cd49d 9f10
Manufactured by BioLegend
APC-conjugated CD49d (9F10) is a flow cytometry reagent used for the detection and analysis of CD49d (very late antigen-4, VLA-4) expression on cells. CD49d is an integrin subunit that is involved in cell-cell and cell-extracellular matrix interactions.
Automatically generated - may contain errors
2 protocols using apc conjugated cd49d 9f10
1
Erythroid Cell Phenotyping by Flow Cytometry
2
Erythroid Cell Phenotyping by Flow Cytometry
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For flow cytometry analysis, in vitro cultured erythroid cells were washed in FACS buffer (1% FBS in PBS) before antibody staining. The following antibodies at indicated dilutions were used in this study: 1:50 APC-conjugated CD235a (Glycophorin A, clone HIR2, 50–153-69, eBioscience), 1:50 FITC-conjugated CD71 (OKT9, 14–0719-82, eBioscience), 1:50 APC-conjugated CD49d (9F10, Biolegend, 304308) and 1:200 FITC-conjugated BAND3 (An et al., 2014 (link); Hu et al., 2013 (link)). For live/ dead cell discrimination Sytox Blue was used according to the manufacturer’s instructions (Thermo Fisher, S34857). For apoptosis analysis, BV421-Annexin V (BD Bioscience, 563973) was used in combination with Sytox Orange according to the manufacturer’s instructions (Thermo Fisher, S34861). FACS analysis was conducted on a BD Bioscience Fortessa flow cytometer at the Whitehead Institute Flow Cytometry core. Data were analyzed using FlowJo software v10.4.2.
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