The Diamond Basophil Isolation Kit has been developed for the two-step isolation of basophilic granulocytes from human PBMCs.

Data and images for Diamond Basophil Isolation Kit, human

Figures

Figure 1

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
Isolated basophils
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Figure 1

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.
View details

Figure 1

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.

Specifications for Diamond Basophil Isolation Kit, human

Overview

The Diamond Basophil Isolation Kit has been developed for the two-step isolation of basophilic granulocytes from human PBMCs.

Detailed product information

Background information

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.

Downstream applications

  • 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

Columns

For the first magnetic separation (depletion): LS or autoMACS
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Columns. For the second magnetic separation (positive selection): MS or autoMACS Columns.

References for Diamond Basophil Isolation Kit, human

Publications

  1. Shiratori, I. et al. (2005) Down-regulation of basophil function by human CD200 and human herpesvirus-8 CD200. J. Immunol. 175: 4441-4449
  2. Sloane et al. (2004) Leukocyte immunoglobulin-like receptors: novel innate receptors for human basophil activation and inhibition. Blood 104: 2832-2839
  3. Falcone et al. (2000) The human basophil: a new appreciation of its role in immune responses. Blood 96: 4028-4038
  4. Karasuyama, H. et al. (2009) Newly discovered roles for basophils: a neglected minority gains new respect. Nat. Rev. Immunol. 9: 9-13

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