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Example of a separation using the REAlease® CD4 (TIL) MicroBead Kit.
A) A human ovarian carcinoma sample was dissociated using the gentleMACS™ Octo Dissociator with Heaters in combination with the Tumor Dissociation Kit, human (# 130-095-929). CD4+
TILs were isolated from the single-cell suspension using the REAlease®
CD4 (TIL) MicroBead Kit, LS Columns, and a MidiMACS™ Separator. Cells were fluorescently stained with CD4-VioGreen™ and analyzed by flow cytometry using the MACSQuant®
Analyzer X. Cell debris and dead cells were excluded from the analysis based on scatter signals and 7-AAD fluorescence. B) The efficient removal of all labels was shown by using Anti-Biotin-APC to analyze the cells by flow cytometry for the presence of REAlease Biotin Complex. Directly after isolation, the cells showed staining of biotin ("MicroBead-free CD4+
cells"), whereas the label-free CD4+
cells after the REAlease Biotin Complex release were negative for biotin similar to the non-labeled cells before separation.
Release efficiency was higher than 99% for the REAlease Anti-Biotin MicroBeads (CD4 (TIL)).
A) Cell purity
B) Label-free cells: REAlease Biotin Complex release
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