Name: REAlease® CD4/CD8 (TIL) MicroBead Kit, human
Availability: InStock

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

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

Documents and Protocols

Brochures/posters

REAlease™ Immunomagnetic Separation Technology - Reversible cell labeling for maximal flexibility

Scientific posters

Tumor-infiltrating T cells: Complete workflows allow faster and improved analysis

Scientific posters

Complete workflows allow comprehensive tumor microenvironment analysis of cell subsets from limited tumor patient samples

App notes/customer reports

Isolation of TILs after tumor dissociation significantly improves the sensitivity of single-cell immune profiling.

Application protocol

Enrichment of bead- and label-free human tumor-infiltrating leukocytes (TILs) from human tumor ...

Data and images