Description
Detailed separation procedure
For depletion, cells are magnetically labeled with an antibody cocktail against so-called lineage markers (CD5, CD45R (B220), CD11b, Gr-1 (Ly-6G/C), 7-4, and Ter-119). In the case of the Lineage Cell Depletion Kit, mouse, the labeling is performed in a two-step procedure. Cells are first labeled with a cocktail of biotinylated antibodies followed by labeling with Anti-Biotin MicroBeads.
In the case of the Direct Lineage Cell Depletion Kit the antibodies are directly coupled to MicroBeads allowing a time-saving one-step labeling and isolation of lineage-negative cells in only 20 minutes.
Downstream applications
The Lineage Cell Depletion Kit has been used for the enrichment of both, hematopoietic stem cells (HSCs)4 and marrow stromal cells (MSCs) from bone marrow5. The labeling procedure leaves lineage-negative cells untouched, allowing further magnetic separation of lineage-negative cells according to expression of markers such as CD1171,2 or Sca-13.
Columns
LS Columns, autoMACS® Columns, or Multi-24 Column Blocks.
Overview
The Direct Lineage Cell Depletion Kit and the indirect Lineage Cell Depletion Kit have been developed for the efficient depletion of mature hematopoietic cells, including T cells, B cells, monocytes/macrophages, granulocytes, erythrocytes and their committed precursors. Both kits are optimized for untouched isolation of lineage-negative cells from mouse bone marrow samples. Using the direct isolation method, depletion can be automated and completed in 20 minutes.
