StraightFrom
^®
Buffy Coat REAlease
^®
CD4 MicroBead Kit
, human

Name: StraightFrom® Buffy Coat REAlease® CD4 MicroBead Kit, human
Availability: InStock

StraightFrom
^®
Buffy Coat REAlease
^®
CD4 MicroBead Kit
, human

The StraightFrom

^®

Buffy Coat CD4 REAlease

^®

MicroBead Kit has been developed for the rapid positive selection of bead-free or label-free CD4

⁺

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

Product data and images for StraightFrom
^®
Buffy Coat REAlease
^®
CD4 MicroBead Kit
, human

Figures

Figure 1

A) CD4

⁺

cells were isolated from a complete buffy coat using the StraightFrom

^®

Buffy Coat REAlease

^®

CD4 MicroBead Kit, Whole Blood Columns, and a MultiMACS™ Cell24 Separator Plus with the Single-Column Adapter. Cells were fluorescently stained with REAfinity™ Antibodies CD4-PE 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 CD4

⁺

cells”), whereas the label-free CD4

⁺

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 CD4 ⁺ cells
View details StraightFrom ^® Buffy Coat REAlease ^® CD4 MicroBead Kit , human Figure 1 A) CD4 ⁺ cells were isolated from a complete buffy coat using the StraightFrom ^® Buffy Coat REAlease ^® CD4 MicroBead Kit, Whole Blood Columns, and a MultiMACS™ Cell24 Separator Plus with the Single-Column Adapter. Cells were fluorescently stained with REAfinity™ Antibodies CD4-PE 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 CD4 ⁺ cells”), whereas the label-free CD4 ⁺ cells after the REAlease Biotin Complex release were negative for biotin similar to the non-labeled cells before separation.	View details StraightFrom ^® Buffy Coat REAlease ^® CD4 MicroBead Kit , human Figure 1 A) CD4 ⁺ cells were isolated from a complete buffy coat using the StraightFrom ^® Buffy Coat REAlease ^® CD4 MicroBead Kit, Whole Blood Columns, and a MultiMACS™ Cell24 Separator Plus with the Single-Column Adapter. Cells were fluorescently stained with REAfinity™ Antibodies CD4-PE 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 CD4 ⁺ cells”), whereas the label-free CD4 ⁺ 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 CD4 ⁺ cells	Label-free CD4 ⁺ cells
View details StraightFrom ^® Buffy Coat REAlease ^® CD4 MicroBead Kit , human Figure 1 A) CD4 ⁺ cells were isolated from a complete buffy coat using the StraightFrom ^® Buffy Coat REAlease ^® CD4 MicroBead Kit, Whole Blood Columns, and a MultiMACS™ Cell24 Separator Plus with the Single-Column Adapter. Cells were fluorescently stained with REAfinity™ Antibodies CD4-PE 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 CD4 ⁺ cells”), whereas the label-free CD4 ⁺ cells after the REAlease Biotin Complex release were negative for biotin similar to the non-labeled cells before separation.	View details StraightFrom ^® Buffy Coat REAlease ^® CD4 MicroBead Kit , human Figure 1 A) CD4 ⁺ cells were isolated from a complete buffy coat using the StraightFrom ^® Buffy Coat REAlease ^® CD4 MicroBead Kit, Whole Blood Columns, and a MultiMACS™ Cell24 Separator Plus with the Single-Column Adapter. Cells were fluorescently stained with REAfinity™ Antibodies CD4-PE 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 CD4 ⁺ cells”), whereas the label-free CD4 ⁺ cells after the REAlease Biotin Complex release were negative for biotin similar to the non-labeled cells before separation.	View details StraightFrom ^® Buffy Coat REAlease ^® CD4 MicroBead Kit , human Figure 1 A) CD4 ⁺ cells were isolated from a complete buffy coat using the StraightFrom ^® Buffy Coat REAlease ^® CD4 MicroBead Kit, Whole Blood Columns, and a MultiMACS™ Cell24 Separator Plus with the Single-Column Adapter. Cells were fluorescently stained with REAfinity™ Antibodies CD4-PE 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 CD4 ⁺ cells”), whereas the label-free CD4 ⁺ cells after the REAlease Biotin Complex release were negative for biotin similar to the non-labeled cells before separation.

Figure

Step-by-step separation procedure

Specifications for StraightFrom
^®
Buffy Coat REAlease
^®
CD4 MicroBead Kit
, human

Overview

The StraightFrom

^®

Buffy Coat CD4 REAlease

^®

MicroBead Kit has been developed for the rapid positive selection of bead-free or label-free CD4

⁺

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

CD4 is an accessory molecule in the recognition of MHC class II/peptide complexes by the TCR heterodimers on CD4

⁺

T helper cells. CD4 is expressed on T helper cells and at a lower level on monocytes and dendritic cells. The CD4 molecule is the receptor for the human immunodeficiency virus. The CD4 antibody recognizes most thymocytes and about 65% of all peripheral blood T cells.

Detailed separation procedure

One entire buffy coat is labeled with StraightFrom

^®

Buffy Coat REAlease

^®

CD4-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 CD4

⁺

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 CD4

⁺

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
^®
CD4 MicroBead Kit
, human

Documents and Protocols

Brochures/posters

StraightFrom® MicroBeads - Cell isolation directly from blood products

Data and images