Name: MACSprep™ CD19 CAR MicroBead Kit, human
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Product data and images for MACSprep™ CD19 CAR MicroBead Kit, human

Figures

Figure 1

**Protocol overview:**
Isolation of CD19 CAR cells from whole blood samples.

Figure 1

Protocol overview:

Isolation of CD19 CAR cells from whole blood samples.

Figure 2

CD19 CAR

⁺

T cells were spiked into a whole blood sample from a healthy donor and isolated using the MACSprep™ CD19 CAR MicroBead Kit, one LS Column, one MS Column, a MACSmix™ Tube Rotator, and a QuadroMACS™ Separator. Cells were fluorescently stained with CD19 CAR Detection Reagent, human, Biotin, Biotin Antibody-PE, CD45-VioBlue

^®

, 7-AAD Staining Solution, CD3-FITC, and analyzed by flow cytometry using the MACSQuant

^®

Analyzer 10. Cell debris and dead cells were excluded from the analysis based on scatter signals and 7-AAD fluorescence. The purity of CD19 CAR

⁺

T cells after separation is defined by the percentage of CD19 CAR

⁺

T cells (viable CD45

^dim/+

, 7-AAD

^–

cells). The purity of CD19 CAR

⁺

T cells varies due to the variation of human samples, experimental set-up, and starting frequency of CD19 CAR

⁺

T cells.

Before separation
View details MACSprep™ CD19 CAR MicroBead Kit, human Figure 2 CD19 CAR ⁺ T cells were spiked into a whole blood sample from a healthy donor and isolated using the MACSprep™ CD19 CAR MicroBead Kit, one LS Column, one MS Column, a MACSmix™ Tube Rotator, and a QuadroMACS™ Separator. Cells were fluorescently stained with CD19 CAR Detection Reagent, human, Biotin, Biotin Antibody-PE, CD45-VioBlue ^® , 7-AAD Staining Solution, CD3-FITC, and analyzed by flow cytometry using the MACSQuant ^® Analyzer 10. Cell debris and dead cells were excluded from the analysis based on scatter signals and 7-AAD fluorescence. The purity of CD19 CAR ⁺ T cells after separation is defined by the percentage of CD19 CAR ⁺ T cells (viable CD45 ^dim/+ , 7-AAD ^– cells). The purity of CD19 CAR ⁺ T cells varies due to the variation of human samples, experimental set-up, and starting frequency of CD19 CAR ⁺ T cells.
After separation with one column	After separation with two columns
View details MACSprep™ CD19 CAR MicroBead Kit, human Figure 2 CD19 CAR ⁺ T cells were spiked into a whole blood sample from a healthy donor and isolated using the MACSprep™ CD19 CAR MicroBead Kit, one LS Column, one MS Column, a MACSmix™ Tube Rotator, and a QuadroMACS™ Separator. Cells were fluorescently stained with CD19 CAR Detection Reagent, human, Biotin, Biotin Antibody-PE, CD45-VioBlue ^® , 7-AAD Staining Solution, CD3-FITC, and analyzed by flow cytometry using the MACSQuant ^® Analyzer 10. Cell debris and dead cells were excluded from the analysis based on scatter signals and 7-AAD fluorescence. The purity of CD19 CAR ⁺ T cells after separation is defined by the percentage of CD19 CAR ⁺ T cells (viable CD45 ^dim/+ , 7-AAD ^– cells). The purity of CD19 CAR ⁺ T cells varies due to the variation of human samples, experimental set-up, and starting frequency of CD19 CAR ⁺ T cells.	View details MACSprep™ CD19 CAR MicroBead Kit, human Figure 2 CD19 CAR ⁺ T cells were spiked into a whole blood sample from a healthy donor and isolated using the MACSprep™ CD19 CAR MicroBead Kit, one LS Column, one MS Column, a MACSmix™ Tube Rotator, and a QuadroMACS™ Separator. Cells were fluorescently stained with CD19 CAR Detection Reagent, human, Biotin, Biotin Antibody-PE, CD45-VioBlue ^® , 7-AAD Staining Solution, CD3-FITC, and analyzed by flow cytometry using the MACSQuant ^® Analyzer 10. Cell debris and dead cells were excluded from the analysis based on scatter signals and 7-AAD fluorescence. The purity of CD19 CAR ⁺ T cells after separation is defined by the percentage of CD19 CAR ⁺ T cells (viable CD45 ^dim/+ , 7-AAD ^– cells). The purity of CD19 CAR ⁺ T cells varies due to the variation of human samples, experimental set-up, and starting frequency of CD19 CAR ⁺ T cells.

Specifications for MACSprep™ CD19 CAR MicroBead Kit, human

Overview

The MACSprep™ CD19 CAR MicroBead Kit, human has been developed for the fast isolation of CD19 CAR cells from freshly drawn anticoagulated whole blood without density gradient centrifugation nor red blood cell (RBC) lysis. During the first isolation step RBCs are aggregated and sedimented. In the second step, CD19 CAR cells are labeled with CD19 CAR Detection Reagent, human, Biotin. After one washing step the CD19 CAR Detection Reagent-bound cells are magnetically labeled with Anti-Biotin MicroBeads. Then, the cell suspension is separated by loading it onto a MACS

^®

Column, which is placed in the magnetic field of a MACS Separator. The magnetically labeled CD19 CAR cells are retained within the column. The unlabeled cells run through, this cell fraction is thus depleted of CD19 CAR cells. After removing the column from the magnetic field, the magnetically retained CD19 CAR cells can be eluted as the positively selected cell fraction.

Detailed product information

Background information

The MACSprep CD19 CAR MicroBead Kit, human has been developed for the separation of cells that are engineered to express CD19-specific chimeric antigen receptor (CAR) on the cell surface.

The kit contains a cocktail and a buffer to remove most of RBCs. In this RBC-reduced blood sample the engineered CD19 CAR T cells can be detected via the CD19 CAR Detection Reagent, human, Biotin and can be enriched by magnetic labeling using Anti-Biotin MicroBeads.

Applications

Isolation of CD19 CAR cells from whole blood. The purified CD19 CAR cells are well suited for further flow cytometric, functional, or molecular analysis.

Resources for MACSprep™ CD19 CAR MicroBead Kit, human

Documents and Protocols

App notes/customer reports

Isolation of CD19 CAR T cells directly from patient samples - Pure sample fraction for further downstream analysis

Data and images