The REAlease
®
CD4/CD8 (TIL) MicroBead Kit,human has been developed for positive selection of CD4
+
and CD8
+
tumor-infiltrating leukocytes (TILs) from single-cell suspensions of solid human tumors as well as xenotransplanted tumors of mice with a humanized immune system. The kit contains an indirect magnetic labeling system for obtaining target cells free of MicroBeads and REAlease Biotin Complex.

Data and images for
REAlease
®
CD4/CD8 (TIL) MicroBead Kit
, human

Figures

Figure 1

Example of a separation using the REAlease® CD4/CD8 (TIL) MicroBead Kit.
A) A human colorectal carcinoma sample was dissociated using the gentleMACS™ Octo Dissociator with Heaters in combination with the Tumor Dissociation Kit, human (# 130-095-929). CD4
+
and CD8
+
TILs were isolated from the single-cell suspension using the REAlease
®
CD4/CD8 (TIL) MicroBead Kit, MS Columns, and a MiniMACS™ Separator. Cells were fluorescently stained with CD3-FITC and analyzed by flow cytometry using the MACSQuant
®
Analyzer 16. 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
+
and CD8
+
cells"), whereas the label-free CD4
+
and CD8
+
cells after the REAlease Biotin Complex release were negative for biotin similar to the non-labeled cells before separation.
Release efficiency was higher than 98.5% for the REAlease Anti-Biotin MicroBeads (CD4/CD8 (TIL)).
A) Cell purity
Before separation
Label-free CD4
+
and CD8
+
cells
View details

Figure 1

Example of a separation using the REAlease® CD4/CD8 (TIL) MicroBead Kit.
A) A human colorectal carcinoma sample was dissociated using the gentleMACS™ Octo Dissociator with Heaters in combination with the Tumor Dissociation Kit, human (# 130-095-929). CD4
+
and CD8
+
TILs were isolated from the single-cell suspension using the REAlease
®
CD4/CD8 (TIL) MicroBead Kit, MS Columns, and a MiniMACS™ Separator. Cells were fluorescently stained with CD3-FITC and analyzed by flow cytometry using the MACSQuant
®
Analyzer 16. 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
+
and CD8
+
cells"), whereas the label-free CD4
+
and CD8
+
cells after the REAlease Biotin Complex release were negative for biotin similar to the non-labeled cells before separation.
Release efficiency was higher than 98.5% for the REAlease Anti-Biotin MicroBeads (CD4/CD8 (TIL)).
View details

Figure 1

Example of a separation using the REAlease® CD4/CD8 (TIL) MicroBead Kit.
A) A human colorectal carcinoma sample was dissociated using the gentleMACS™ Octo Dissociator with Heaters in combination with the Tumor Dissociation Kit, human (# 130-095-929). CD4
+
and CD8
+
TILs were isolated from the single-cell suspension using the REAlease
®
CD4/CD8 (TIL) MicroBead Kit, MS Columns, and a MiniMACS™ Separator. Cells were fluorescently stained with CD3-FITC and analyzed by flow cytometry using the MACSQuant
®
Analyzer 16. 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
+
and CD8
+
cells"), whereas the label-free CD4
+
and CD8
+
cells after the REAlease Biotin Complex release were negative for biotin similar to the non-labeled cells before separation.
Release efficiency was higher than 98.5% for the REAlease Anti-Biotin MicroBeads (CD4/CD8 (TIL)).
B) Label-free cells: REAlease Biotin Complex release
Before separation
MicroBead-free CD4
+
and CD8
+
cells
Label-free CD4
+
and CD8
+
cells
View details

Figure 1

Example of a separation using the REAlease® CD4/CD8 (TIL) MicroBead Kit.
A) A human colorectal carcinoma sample was dissociated using the gentleMACS™ Octo Dissociator with Heaters in combination with the Tumor Dissociation Kit, human (# 130-095-929). CD4
+
and CD8
+
TILs were isolated from the single-cell suspension using the REAlease
®
CD4/CD8 (TIL) MicroBead Kit, MS Columns, and a MiniMACS™ Separator. Cells were fluorescently stained with CD3-FITC and analyzed by flow cytometry using the MACSQuant
®
Analyzer 16. 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
+
and CD8
+
cells"), whereas the label-free CD4
+
and CD8
+
cells after the REAlease Biotin Complex release were negative for biotin similar to the non-labeled cells before separation.
Release efficiency was higher than 98.5% for the REAlease Anti-Biotin MicroBeads (CD4/CD8 (TIL)).
View details

Figure 1

Example of a separation using the REAlease® CD4/CD8 (TIL) MicroBead Kit.
A) A human colorectal carcinoma sample was dissociated using the gentleMACS™ Octo Dissociator with Heaters in combination with the Tumor Dissociation Kit, human (# 130-095-929). CD4
+
and CD8
+
TILs were isolated from the single-cell suspension using the REAlease
®
CD4/CD8 (TIL) MicroBead Kit, MS Columns, and a MiniMACS™ Separator. Cells were fluorescently stained with CD3-FITC and analyzed by flow cytometry using the MACSQuant
®
Analyzer 16. 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
+
and CD8
+
cells"), whereas the label-free CD4
+
and CD8
+
cells after the REAlease Biotin Complex release were negative for biotin similar to the non-labeled cells before separation.
Release efficiency was higher than 98.5% for the REAlease Anti-Biotin MicroBeads (CD4/CD8 (TIL)).
View details

Figure 1

Example of a separation using the REAlease® CD4/CD8 (TIL) MicroBead Kit.
A) A human colorectal carcinoma sample was dissociated using the gentleMACS™ Octo Dissociator with Heaters in combination with the Tumor Dissociation Kit, human (# 130-095-929). CD4
+
and CD8
+
TILs were isolated from the single-cell suspension using the REAlease
®
CD4/CD8 (TIL) MicroBead Kit, MS Columns, and a MiniMACS™ Separator. Cells were fluorescently stained with CD3-FITC and analyzed by flow cytometry using the MACSQuant
®
Analyzer 16. 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
+
and CD8
+
cells"), whereas the label-free CD4
+
and CD8
+
cells after the REAlease Biotin Complex release were negative for biotin similar to the non-labeled cells before separation.
Release efficiency was higher than 98.5% for the REAlease Anti-Biotin MicroBeads (CD4/CD8 (TIL)).

Specifications for
REAlease
®
CD4/CD8 (TIL) MicroBead Kit
, human

Overview

The REAlease
®
CD4/CD8 (TIL) MicroBead Kit,human has been developed for positive selection of CD4
+
and CD8
+
tumor-infiltrating leukocytes (TILs) from single-cell suspensions of solid human tumors as well as xenotransplanted tumors of mice with a humanized immune system. The kit contains an indirect magnetic labeling system for obtaining target cells free of MicroBeads and REAlease Biotin Complex.

Detailed product information

Background information

The REAlease
®
CD4/CD8 (TIL) MicroBead Kit has been developed for the isolation of CD4
+
and CD8
+
tumor-infiltrating leukocytes (TILs) from single-cell suspensions of solid human tumors as well as xenotransplanted tumors of mice with a humanized immune system. CD4 is expressed on T helper cells, at a lower level on monocytes and dendritic cells, and at a very low level on hematopoietic progenitor cells. CD8 is expressed on cytotoxic T cells.
The REAlease CD4/CD8 (TIL) MicroBead Kit is an indirect magnetic labeling system that allows to obtain cells free of MicroBeads and the REAlease Biotin Complex.

Downstream applications

  • Isolation of specific CD4+ and CD8+ tumor-infiltrating T cell subsets. The selected CD4+ and CD8+ T cell populations can be sorted according to a second marker of interest, for example, CD137 (4-1BB), CD223 (LAG-3), CD279 (PD1), or CD366 (TIM-3).
  • Isolation of CD4+ and CD8+ tumor-infiltrating T cells which need to be label-free.

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REAlease
®
CD4/CD8 (TIL) MicroBead Kit
, human

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