A375 cells stably expressing enCas12a were transduced with 6 triple knockout arrays, 3 single knockout constructs, and one empty control vector. Two days after transduction, cells were selected with puromycin (1μg/mL), and selected on puromycin for 7 days. Cells were visualized by flow cytometry on the BDAccuri C6 Sampler. To prepare samples for visualization, cells were stained with FITC anti-human CD47 (Biolegend # 323106), PE anti-human CD63 (Biolegend #353004) and APC anti-human β2-microglobulin (Biolegend #316312) antibodies, diluted 1:100 in flow buffer (PBS, 2% FBS, 5μM EDTA), incubated for 30 min on ice, washed with flow buffer twice to remove residual antibody, and resuspended in flow buffer. Flow cytometry data were analyzed using FlowJo (v10). Gates were set such that ~1% of cells score as knockout in the control condition. Compensation was applied using single stained empty vector control and triple stained empty vector control cell populations. Zero, single, double, and triple knockout populations were quantified using boolean gating in FlowJo.
Apc anti human β2 microglobulin
Manufactured by BioLegend
APC anti-human β2-microglobulin is a fluorescently-labeled antibody that binds to the β2-microglobulin protein found on the surface of human cells. It can be used for the detection and analysis of β2-microglobulin-expressing cells in various applications, such as flow cytometry.
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Lab products found in correlation
2 protocols using apc anti human β2 microglobulin
1
Multiplexed Knockout Screening in A375 Cells
2
Multiplexed Knockout Screening in A375 Cells
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A375 cells stably expressing enCas12a were transduced with 6 triple knockout arrays, 3 single knockout constructs, and one empty control vector. Two days after transduction, cells were selected with puromycin (1μg/mL), and selected on puromycin for 7 days. Cells were visualized by flow cytometry on the BDAccuri C6 Sampler. To prepare samples for visualization, cells were stained with FITC anti-human CD47 (Biolegend # 323106), PE anti-human CD63 (Biolegend #353004) and APC anti-human β2-microglobulin (Biolegend #316312) antibodies, diluted 1:100 in flow buffer (PBS, 2% FBS, 5μM EDTA), incubated for 30 min on ice, washed with flow buffer twice to remove residual antibody, and resuspended in flow buffer. Flow cytometry data were analyzed using FlowJo (v10). Gates were set such that ~1% of cells score as knockout in the control condition. Compensation was applied using single stained empty vector control and triple stained empty vector control cell populations. Zero, single, double, and triple knockout populations were quantified using boolean gating in FlowJo.
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