YILIMART - Basophil Cell Isolation Kit II, human

The Basophil Isolation Kit II is an indirect magnetic labeling system for the negative isolation of basophils from PBMCs.

Detailed separation procedure

Non-basophils (i.e. T cells, NK cells, B cells, monocytes, dendritic cells, erythroid cells, platelets, neutrophils, and eosinophils) are indirectly magnetically labeled using a cocktail of biotin-conjugated antibodies and Anti-Biotin MicroBeads. Isolation of highly pure basophils is achieved by depletion of the magnetically labeled cells.

Downstream applications

Isolated basophils have been used for phenotypical characterization

³

, for histamine, leukotriene, and cytokine

^{1, 4, 5, 6}

release assays, for molecular studies

^{1, 2, 5}

, protein analysis

^{4, 5}

, and other applications.

Columns

LS, XS, or autoMACS

^®

Columns.

Figure 1

Basophils were enriched from human PBMCs using the Basophil Isolation Kit II, a MidiMACS™ Separator and an LS Column. Cells were stained with CD203c-APC, CD123-PE and CD303 (BDCA-2)-FITC. Basophils were detected as CD203c

⁺

cells or CD123

^bright

CD303

^–

cells because CD123 is expressed at high levels on both basophils and CD303

⁺

plasmacytoid dendritic cells.

Unseparated fraction
A:	B:

Isolated basophils
A:	B:

Figure 2
View details

Figure 2

Basophils isolated with MACS
^®
Technology using the Basophil Isolation Kit II. (Courtesy of Dr. Karin Haisch, Borstel, Germany.)

Basophils isolated with MACS
^®
Technology using the Basophil Isolation Kit II. (Courtesy of Dr. Karin Haisch, Borstel, Germany.)

Selected references
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. Nishiyama, C. et al. (2004) Polymorphisms in the Fc epsilon RI beta promoter region affecting transcription activity: a possible promoter-dependent mechanism for association between Fc epsilon RI beta and atopy. J. Immunol. 173: 6458-6464
3. de Paulis, A. et al. (2004) Urokinase induces basophil chemotaxis through a urokinase receptor epitope that is an endogenous ligand for formyl peptide receptor-like 1 and -like 2. J. Immunol. 173: 5739-5748
4. Gibbs, B. F. et al. (2002) Regulation of mediator secretion in human basophils by p38 mitogen-activated protein kinase: phosphorylation is sensitive to the effects of phosphatidylinositol 3-kinase inhibitors and calcium mobilization. J. Leukoc. Biol. 72: 391-400
5. Lavens-Phillips, S.E. et al. (2000) The tyrosine kinases p53/56lyn and p72syk are differentially expressed at the protein level but not at the messenger RNA level in nonreleasing human basophils. Am. J. Respir. Cell Mol. Biol. 23: 566-571
6. Haisch, K. et al. (1999) Purification of morphologically and functionally intact human basophils to near homogeneity. J. Immunol. Methods 226: 129-137

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Basophil Cell Isolation Kit II, human

For research use only

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Detailed separation procedure

Downstream applications

Columns

Figure 1

Figure 2

Figure 2

Selected references

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