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Methods Mol Biol. 2010;647:377-90. doi: 10.1007/978-1-60761-738-9_23.

Flow cytometry analysis of transcription factors in T lymphocytes.

Author information

1
Center for Cell Biology and Cancer Research, Albany Medical College, Albany, NY, USA.

Abstract

Detection of transcription factors in immune cell populations, particularly in subpopulations that are represented at low frequencies in lymphoid and nonlymphoid organs, presents a particular challenge when using traditional methods such as western blot analysis. Therefore, development of flow cytometry-based methods which allow identification of transcription factors in specific immune cell populations is of main interest. Here we developed and optimized a methodology for rapid and convenient detection of the transcription factor BCL11B in T lymphocyte subpopulations using flow cytometry. The optimal protocol employs saponin and Tween 20 both during the fixation and permeabilization steps, and we demonstrate that it is efficient for three anti-BCL11B antibodies covering distinctive BCL11B epitopes. In addition, we prove that the method preserves the staining of surface markers.

PMID:
20694680
PMCID:
PMC3788577
DOI:
10.1007/978-1-60761-738-9_23
[Indexed for MEDLINE]
Free PMC Article

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