The REAlease
®
CD62L MicroBead Kit has been developed for positive selection of CD62L
+
cells from PBMCs. The kit contains an indirect magnetic labeling system for obtaining target cells free of MicroBeads and REAlease Biotin Complex.

Data and images for
REAlease
®
CD62L MicroBead Kit
, human

Figures

Figure 1

View details
Step-by-step REAlease MicroBeads separation procedure

Figure 1

Step-by-step REAlease MicroBeads separation procedure

Figure 2

Example of a separation using the REAlease® CD62L MicroBead Kit
A) CD62L
+
cells were isolated from human PBMCs using the REAlease
®
CD62L MicroBead Kit, MS Columns, and a MiniMACS™ Separator. Cells were fluorescently stained with CD62L-PE and CD4-PE-Vio
®
770 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 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 CD62L
+
cells”), whereas the label-free CD62L
+
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 CD62L
+
cells
View details

Figure 2

Example of a separation using the REAlease® CD62L MicroBead Kit
A) CD62L
+
cells were isolated from human PBMCs using the REAlease
®
CD62L MicroBead Kit, MS Columns, and a MiniMACS™ Separator. Cells were fluorescently stained with CD62L-PE and CD4-PE-Vio
®
770 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 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 CD62L
+
cells”), whereas the label-free CD62L
+
cells after the REAlease Biotin Complex release were negative for biotin similar to the non-labeled cells before separation.
View details

Figure 2

Example of a separation using the REAlease® CD62L MicroBead Kit
A) CD62L
+
cells were isolated from human PBMCs using the REAlease
®
CD62L MicroBead Kit, MS Columns, and a MiniMACS™ Separator. Cells were fluorescently stained with CD62L-PE and CD4-PE-Vio
®
770 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 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 CD62L
+
cells”), whereas the label-free CD62L
+
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 CD62L
+
cells
Label-free CD62L
+
cells
View details

Figure 2

Example of a separation using the REAlease® CD62L MicroBead Kit
A) CD62L
+
cells were isolated from human PBMCs using the REAlease
®
CD62L MicroBead Kit, MS Columns, and a MiniMACS™ Separator. Cells were fluorescently stained with CD62L-PE and CD4-PE-Vio
®
770 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 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 CD62L
+
cells”), whereas the label-free CD62L
+
cells after the REAlease Biotin Complex release were negative for biotin similar to the non-labeled cells before separation.
View details

Figure 2

Example of a separation using the REAlease® CD62L MicroBead Kit
A) CD62L
+
cells were isolated from human PBMCs using the REAlease
®
CD62L MicroBead Kit, MS Columns, and a MiniMACS™ Separator. Cells were fluorescently stained with CD62L-PE and CD4-PE-Vio
®
770 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 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 CD62L
+
cells”), whereas the label-free CD62L
+
cells after the REAlease Biotin Complex release were negative for biotin similar to the non-labeled cells before separation.
View details

Figure 2

Example of a separation using the REAlease® CD62L MicroBead Kit
A) CD62L
+
cells were isolated from human PBMCs using the REAlease
®
CD62L MicroBead Kit, MS Columns, and a MiniMACS™ Separator. Cells were fluorescently stained with CD62L-PE and CD4-PE-Vio
®
770 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 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 CD62L
+
cells”), whereas the label-free CD62L
+
cells after the REAlease Biotin Complex release were negative for biotin similar to the non-labeled cells before separation.

Specifications for
REAlease
®
CD62L MicroBead Kit
, human

Overview

The REAlease
®
CD62L MicroBead Kit has been developed for positive selection of CD62L
+
cells from PBMCs. 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 CD62L antigen is a 74 kDa glycoprotein and is a member of the selectin family of cell surface molecules, also referred to as L-selectin, LECAM-1, or LAM-1. CD62L binds a series of glycoproteins including CD34, GlyCAM-1, and MAdCAM-1. CD62L is important for homing of naive lymphocytes via the high endothelial venules to peripheral lymph nodes and Peyer's patches. The CD62L antigen also contributes to the recruitment of leukocytes from the blood to areas of inflammation. Most hematopoietic cells, including most peripheral blood B cells, T cells, monocytes, dendritic cells, granulocytes, and some myeloid cells from bone marrow and thymocytes, express CD62L. CD62L is continuously endoproteolytically cleaved from the cell surface of CD62L-expressing neutrophils and lymphocytes (shedding). Proteolysis is accelerated, e.g., after antigen activation of T cells.

Downstream applications

  • Isolation of label-free CD62L+, CD3+, CD4+, and CD8+ T cells using first the REAlease® CD62L MicroBead Kit, human, followed by a second enrichment using REAlease CD3, CD4, or CD8 MicroBead Kits, human.
  • Isolation of central memory T cells by using the REAlease CD62L MicroBead Kit, human, followed by cell enrichment using CD45RO MicroBeads, human (#130-046-001). For further subset discrimination this isolation strategy may be combined with REAlease CD3, CD4, or CD8 MicroBead Kits, human.
  • Positive selection of label-free CD62L+ T cells from pre-enriched CD4+ or CD8+ T cells using the CD4+ T Cell Isolation Kit (# 130-096-533) or the CD8+ T Cell Isolation Kit (# 130-096-495).
  • Isolation of label-free CD4+CD62L+ central memory T cells from pre-enriched CD4+ memory T cells using the Memory CD4+ T Cell Isolation Kit (# 130-091-893).

Columns

MS or LS Columns.

Resources for
REAlease
®
CD62L MicroBead Kit
, human

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®
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