CAR (Whitlow/218 Linker) Cell Enrichment Kit #65686
Product Information
Protocol
Product Description
The CAR (Whitlow/218 Linker) Cell Enrichment Kit is designed to positively select cells that are engineered to express a Whitlow/218 linker-containing single-chain variable fragment (scFv)-based chimeric antigen receptor (CAR) on the cell surface. Using an indirect magnetic bead-based immunoaffinity enrichment protocol, this kit yields highly pure and viable CAR positive cells that are bead-free. Within a heterogeneous population of cells, desired CAR expressing cells are labeled with a biotinylated rabbit monoclonal antibody targeting the Whitlow/218 linker. Releasable Streptavidin Beads are then incubated with the cell mixture. Bead-bound cells are isolated without columns using a magnet (not included), and then beads are removed from cells using DNase I. Following release from beads, the positively selected fraction of CAR positive cells containing surface-bound biotinylated antibody-streptavidin complexes are available for expansion or use in downstream applications such as flow cytometry and gene expression analyses.
Bead Specifications:
Releasable Streptavidin Beads are paramagnetic polystyrene beads (4.5 μm diameter) coupled to streptavidin via a DNA linker susceptible to cleavage by DNase I. The beads provided in this kit are not recommended for the isolation of phagocytic cells as the beads are likely to be engulfed by these cells.
Bead Specifications:
Releasable Streptavidin Beads are paramagnetic polystyrene beads (4.5 μm diameter) coupled to streptavidin via a DNA linker susceptible to cleavage by DNase I. The beads provided in this kit are not recommended for the isolation of phagocytic cells as the beads are likely to be engulfed by these cells.
Specificity / Sensitivity
Biotinylated Whitlow/218 Linker (E3U7Q) Rabbit mAb recognizes exogenously expressed levels of scFv-based CARs containing a Whitlow/218 linker.
Species Reactivity:
All Species Expected
Background
Magnetic bead-based immunoaffinity cell enrichment is a gentle purification method that can be leveraged, in part, to facilitate a robust interrogation of the biology of immune cells that are engineered to express CARs. For example, isolation of rare subsets of CAR positive cells from a complex, heterogeneous population of cells presents the opportunity for extensive downstream analyses using single-cell omics assays. In the context of CAR cell engineering, magnetic bead-based immunoaffinity cell enrichment can also be useful in scenarios when delivery of the CAR transgene is inefficient, resulting in a small population of cells expressing the CAR transgene.
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For Research Use Only. Not For Use In Diagnostic Procedures.
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