The StraightFrom
®
Buffy Coat CD3 REAlease
®
MicroBead Kit has been developed for the rapid positive selection of bead-free or label-free CD3
+
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
®
CD3 MicroBead Kit
, human

Figures

Figure 1

A) CD3
+
cells were isolated from a complete buffy coat using the StraightFrom
®
Buffy Coat REAlease
®
CD3 MicroBead Kit, Whole Blood Columns, and a MultiMACS™ Cell24 Separator Plus with the Single-Column Adapter. Cells were fluorescently stained with REAfinity™ Antibodies CD3-FITC, CD8-APC, and 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 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 CD3
+
cells”), whereas the label-free CD3
+
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 CD3
+
cells
View details

Figure 1

A) CD3
+
cells were isolated from a complete buffy coat using the StraightFrom
®
Buffy Coat REAlease
®
CD3 MicroBead Kit, Whole Blood Columns, and a MultiMACS™ Cell24 Separator Plus with the Single-Column Adapter. Cells were fluorescently stained with REAfinity™ Antibodies CD3-FITC, CD8-APC, and 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 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 CD3
+
cells”), whereas the label-free CD3
+
cells after the REAlease Biotin Complex release were negative for biotin similar to the non-labeled cells before separation.
View details

Figure 1

A) CD3
+
cells were isolated from a complete buffy coat using the StraightFrom
®
Buffy Coat REAlease
®
CD3 MicroBead Kit, Whole Blood Columns, and a MultiMACS™ Cell24 Separator Plus with the Single-Column Adapter. Cells were fluorescently stained with REAfinity™ Antibodies CD3-FITC, CD8-APC, and 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 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 CD3
+
cells”), whereas the label-free CD3
+
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 CD3
+
cells
Label-free CD3
+
cells
View details

Figure 1

A) CD3
+
cells were isolated from a complete buffy coat using the StraightFrom
®
Buffy Coat REAlease
®
CD3 MicroBead Kit, Whole Blood Columns, and a MultiMACS™ Cell24 Separator Plus with the Single-Column Adapter. Cells were fluorescently stained with REAfinity™ Antibodies CD3-FITC, CD8-APC, and 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 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 CD3
+
cells”), whereas the label-free CD3
+
cells after the REAlease Biotin Complex release were negative for biotin similar to the non-labeled cells before separation.
View details

Figure 1

A) CD3
+
cells were isolated from a complete buffy coat using the StraightFrom
®
Buffy Coat REAlease
®
CD3 MicroBead Kit, Whole Blood Columns, and a MultiMACS™ Cell24 Separator Plus with the Single-Column Adapter. Cells were fluorescently stained with REAfinity™ Antibodies CD3-FITC, CD8-APC, and 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 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 CD3
+
cells”), whereas the label-free CD3
+
cells after the REAlease Biotin Complex release were negative for biotin similar to the non-labeled cells before separation.
View details

Figure 1

A) CD3
+
cells were isolated from a complete buffy coat using the StraightFrom
®
Buffy Coat REAlease
®
CD3 MicroBead Kit, Whole Blood Columns, and a MultiMACS™ Cell24 Separator Plus with the Single-Column Adapter. Cells were fluorescently stained with REAfinity™ Antibodies CD3-FITC, CD8-APC, and 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 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 CD3
+
cells”), whereas the label-free CD3
+
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
®
CD3 MicroBead Kit
, human

Overview

The StraightFrom
®
Buffy Coat CD3 REAlease
®
MicroBead Kit has been developed for the rapid positive selection of bead-free or label-free CD3
+
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

CD3 is expressed on all T cells and CD56
+
NK T cells and is associated with the T cell receptor (TCR) heterodimer. Most of the human peripheral blood lymphocytes (70–80%) and thymocytes (65–85%) are CD3
+
. The epitope recognized by the REAlease
®
CD3-Biotin Complex is located on the CD3ε chain.

Detailed separation procedure

One entire buffy coat is labeled with StraightFrom
®
Buffy Coat REAlease
®
CD3-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 CD3
+
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 CD3
+
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
®
CD3 MicroBead Kit
, human