At day 0, CD34+ HSPCs were thawed and stained with the following antibody panel: CD34 BV 421, CD38 APC-Cy7 (clone HIT2, BioLegend), CD90 PE-Cy7 (clone 5E10, BioLegend) and CD45RA APC (clone HI100, BioLegend). The cells were then FACS-sorted into different subsets including CD38+ cells, haematopoietic stem cells (HSCs) and multipotent progenitors (MPPs). Unsorted cells were taken as an experimental control. After 2 days, both sorted and unsorted cells were electroporated and transduced with AAV6_PGK-GFP. At days 5 and 12 post editing, the cells were stained with the above antibody panels to determine the percentage of HSCs and MPPs as well as evaluate GFP expression by FACS. Cell yields were calculated by adding 10 microliters of Count Bright Absolute Counting Beads (ThermoFisher Scientific), following the manufacturer’s protocol.
Cd90 pe cy7 clone 5e10
Manufactured by BioLegend
CD90 PE-Cy7 (clone 5E10) is a fluorescently labeled monoclonal antibody that binds to the CD90 antigen. CD90 is a glycosylphosphatidylinositol (GPI)-anchored cell surface protein expressed on a variety of cell types, including hematopoietic stem cells, mesenchymal stem cells, and certain T cell subsets.
Automatically generated - may contain errors
2 protocols using cd90 pe cy7 clone 5e10
1
Purification and Genetic Modification of HSPCs
2
Tracking Edited Hematopoietic Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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At day 0, CD34 + HSPCs were thawed and stained with the following antibody panel: CD34 BV 421, CD38 APC-Cy7 (clone HIT2, BioLegend), CD90 PE-Cy7 (clone 5E10, BioLegend) and CD45RA APC (clone HI100, BioLegend). The cells were then FACS-sorted into different subsets including CD38+ cells, haematopoietic stem cells (HSCs) and multipotent progenitors (MPPs). Unsorted cells were taken as an experimental control. After 2 days, both sorted and unsorted cells were electroporated and transduced with AAV6_PGK-GFP. At days 5 and 12 post editing, the cells were stained with the above antibody panels to determine the percentage of HSCs and MPPs as well as evaluate GFP expression by FACS. Cell yields were calculated by adding 10 microliters of Count Bright Absolute Counting Beads (ThermoFisher Scientific), following the manufacturer's protocol.
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