The StraightFrom
®
Buffy Coat CD8 REAlease
®
MicroBead Kit has been developed for the rapid positive selection of bead-free or label-free CD8
+
cells, starting directly from one buffy coat using the MultiMACS™ Cell24 Separator Plus for semi-automated separation or a MACS
®
Separator for manual separation. No sample preparation is required, including density gradient centrifugation, erythrocyte lysis, wash steps, or cell count. Whole Blood Columns are included in the kit to allow

Data and images for
StraightFrom
®
Buffy Coat REAlease
®
CD8 MicroBead Kit
, human

Figures

Figure 1

A) CD8
+
cells were isolated from a complete buffy coat using the StraightFrom
®
Buffy Coat REAlease
®
CD8 MicroBead Kit, Whole Blood Columns, and a MultiMACS™ Cell24 Separator Plus with the Single-Column Adapter. Cells were fluorescently stained with REAfinity™ Antibodies CD8-APC 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 propidium iodide fluorescence.
B) The efficient removal of all labels was shown by using Biotin-APC-Vio
®
770 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 CD8
+
cells”), whereas the label-free CD8
+
cells after the REAlease Biotin Complex release were negative for biotin similar to the non-labeled cells before separation.
A) Cell purity
Before separation
Label-free CD8
+
cells
View details

Figure 1

A) CD8
+
cells were isolated from a complete buffy coat using the StraightFrom
®
Buffy Coat REAlease
®
CD8 MicroBead Kit, Whole Blood Columns, and a MultiMACS™ Cell24 Separator Plus with the Single-Column Adapter. Cells were fluorescently stained with REAfinity™ Antibodies CD8-APC 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 propidium iodide fluorescence.
B) The efficient removal of all labels was shown by using Biotin-APC-Vio
®
770 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 CD8
+
cells”), whereas the label-free CD8
+
cells after the REAlease Biotin Complex release were negative for biotin similar to the non-labeled cells before separation.
View details

Figure 1

A) CD8
+
cells were isolated from a complete buffy coat using the StraightFrom
®
Buffy Coat REAlease
®
CD8 MicroBead Kit, Whole Blood Columns, and a MultiMACS™ Cell24 Separator Plus with the Single-Column Adapter. Cells were fluorescently stained with REAfinity™ Antibodies CD8-APC 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 propidium iodide fluorescence.
B) The efficient removal of all labels was shown by using Biotin-APC-Vio
®
770 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 CD8
+
cells”), whereas the label-free CD8
+
cells after the REAlease Biotin Complex release were negative for biotin similar to the non-labeled cells before separation.
B) Label-free cells: REAlease Biotin Complex release
Before separation
MicroBead-free CD8
+
cells
Label-free CD8
+
cells
View details

Figure 1

A) CD8
+
cells were isolated from a complete buffy coat using the StraightFrom
®
Buffy Coat REAlease
®
CD8 MicroBead Kit, Whole Blood Columns, and a MultiMACS™ Cell24 Separator Plus with the Single-Column Adapter. Cells were fluorescently stained with REAfinity™ Antibodies CD8-APC 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 propidium iodide fluorescence.
B) The efficient removal of all labels was shown by using Biotin-APC-Vio
®
770 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 CD8
+
cells”), whereas the label-free CD8
+
cells after the REAlease Biotin Complex release were negative for biotin similar to the non-labeled cells before separation.
View details

Figure 1

A) CD8
+
cells were isolated from a complete buffy coat using the StraightFrom
®
Buffy Coat REAlease
®
CD8 MicroBead Kit, Whole Blood Columns, and a MultiMACS™ Cell24 Separator Plus with the Single-Column Adapter. Cells were fluorescently stained with REAfinity™ Antibodies CD8-APC 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 propidium iodide fluorescence.
B) The efficient removal of all labels was shown by using Biotin-APC-Vio
®
770 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 CD8
+
cells”), whereas the label-free CD8
+
cells after the REAlease Biotin Complex release were negative for biotin similar to the non-labeled cells before separation.
View details

Figure 1

A) CD8
+
cells were isolated from a complete buffy coat using the StraightFrom
®
Buffy Coat REAlease
®
CD8 MicroBead Kit, Whole Blood Columns, and a MultiMACS™ Cell24 Separator Plus with the Single-Column Adapter. Cells were fluorescently stained with REAfinity™ Antibodies CD8-APC 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 propidium iodide fluorescence.
B) The efficient removal of all labels was shown by using Biotin-APC-Vio
®
770 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 CD8
+
cells”), whereas the label-free CD8
+
cells after the REAlease Biotin Complex release were negative for biotin similar to the non-labeled cells before separation.

Figure

View details
Step-by-step separation procedure
Step-by-step separation procedure

Specifications for
StraightFrom
®
Buffy Coat REAlease
®
CD8 MicroBead Kit
, human

Overview

The StraightFrom
®
Buffy Coat CD8 REAlease
®
MicroBead Kit has been developed for the rapid positive selection of bead-free or label-free CD8
+
cells, starting directly from one buffy coat using the MultiMACS™ Cell24 Separator Plus for semi-automated separation or a MACS
®
Separator for manual separation. No sample preparation is required, including density gradient centrifugation, erythrocyte lysis, wash steps, or cell count. Whole Blood Columns are included in the kit to allow direct processing of an entire buffy coat with a simple and quick protocol.

Detailed product information

Background information

The CD8 antigen forms a complex together with the T cell receptor and acts as an accessory molecule in the recognition of MHC class I/peptide complexes by the TCR heterodimer on CD8
+
cytotoxic T cells. The CD8 molecule consists of either an α/β heterodimer or an α/α homodimer. It is expressed strongly on cytotoxic T cells and dimly on a subset of NK cells. CD8 is found on most thymocytes and on about one third of all peripheral blood T cells. CD8
+
cytotoxic T cells play an important role in the killing of virus-infected cells and tumor cells.

Detailed separation procedure

One entire buffy coat is labeled with StraightFrom
®
Buffy Coat REAlease
®
CD8-Biotin Complex followed by Anti-Biotin MicroBeads and loaded onto Whole Blood Columns, which are placed in the magnetic field of a MultiMACS™ Cell24 Separator Plus. After removing the column from the magnetic field, the magnetically retained CD8
+
cells are eluted as the positively selected cell fraction bead-free, or free of MicroBeads. Optionally, the REAlease Biotin Complex can be removed for label-free target cells.

Downstream applications

The purified CD8
+
cells are bead-free, practical for consecutive magnetic isolation and optionally label-free with epitopes available for any downstream application. The isolated cells are suited for further flow cytometric analyses, molecular biology applications, and functional studies.

Resources for
StraightFrom
®
Buffy Coat REAlease
®
CD8 MicroBead Kit
, human