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Example of a separation using the REAlease ® CD45 (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). CD45+
TILs were isolated from the single-cell suspension using the REAlease®
CD45 (TIL) MicroBead Kit, LS Columns, and a MidiMACS™ Separator. Cells were fluorescently stained with CD45-VioBlue®
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 CD45+
cells"), whereas the label-free CD45+
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 (CD45 (TIL)).
A) Cell purity
B) Label-free cells: REAlease Biotin Complex release
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