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, HSCs and multipotent progenitors (MPPs). Unsorted cells were used as an additional cell population. 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 flow cytometry. Cell yields were calculated by adding 10 μL of Count Bright Absolute Counting Beads (ThermoFisher Scientific), following the manufacturer’s protocol.
Cd38 apc cy7 clone hit2
Manufactured by BioLegend
CD38 APC-Cy7 (clone HIT2) is a fluorescently-labeled antibody that binds to the CD38 antigen expressed on the surface of various cell types. It can be used for the detection and analysis of CD38-positive cells in flow cytometry applications.
Automatically generated - may contain errors
3 protocols using cd38 apc cy7 clone hit2
1
Editing Hematopoietic Stem Cells
2
Purification and Genetic Modification of HSPCs
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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.
3
Tracking Edited Hematopoietic Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
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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