The Diamond Basophil Isolation Kit has been developed for the two-step isolation of basophilic granulocytes from human PBMCs.
Basophilic granulocytes, also known as basophils, are a rare population of leukocytes with frequencies below 1% in peripheral blood. First described by Paul Ehrlich in 1879, basophils have long been considered as circulating mast cells. They express high-affinity immunoglobulin (Ig) E receptors (FcεRI). Upon receptor cross-linking by binding of antigen-IgE complexes, basophils degranulate and thereby release inflammatory mediators such as histamine and leukotriene. Thus, basophils play a major role in allergy, asthma, and immunity.1-4
Detailed separation procedure
First, non-basophils are magnetically labeled for depletion over a MACS Column. The pre-enriched non-labeled basophils in the flow-through are then positively selected via CD123 MicroBeads.
- Studies on signal requirements for basophil activation, induction of mediator synthesis and release etc.
- Studies on the involvement of basophils in hypersensitivity reactions
- Studies on cytokine expression by basophils
For the first magnetic separation (depletion): LS or autoMACS® Columns. For the second magnetic separation (positive selection): MS or autoMACS Columns.
Basophils were isolated from human PBMCs using the Diamond Basophil Isolation Kit and two MS Columns. Cells were fluorescently stained with CD123-PE and CD203c-APC and analyzed by flow cytometry. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.
PBMCs before separation