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Detection of True IgE-expressing Mouse B Lineage Cells
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Immunology and Infection
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JoVE Journal Immunology and Infection
Detection of True IgE-expressing Mouse B Lineage Cells

Detection of True IgE-expressing Mouse B Lineage Cells

DOI:
10.3791/52264-v

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09:40 min

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December 01, 2014

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, , ,
1Department of Medicine, Division of Rheumatology, Immunology and Allergy,Brigham and Women’s Hospital and Harvard Medical School

Chapters

  • 00:05Title
  • 01:54Purification and Activation of B Cells
  • 03:32Trypsinization, Fixation and Permeabilization
  • 06:23Flourescence Staining for IgG1 and IgE Class Switching
  • 08:37Results: Improvements Seen with Trypsinization
  • 09:11Conclusion

Summary

Automatic Translation

Automatic Translation

In vitro analysis of class switch recombination in mice is challenging due to cytophilic IgE molecules bound to Fc receptors on the surface of B cells. We describe a method for IgE detection using trypsin-mediated cleavage of surface-bound IgE prior to fixation and permeabilization for cytoplasmic fluorescence staining.

Tags

Keywords: IgEB LymphocyteClass Switch Recombination (CSR)Fc&949;RII/CD23Flow CytometryTrypsin-mediated CleavageCytoplasmic StainingIn Vitro AssayMolecular And Cellular Immunology

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