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.
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Abbreviations
- BCL11B:
-
B-cell leukemia/lymphoma
- CTIP2:
-
Chicken ovalbumin upstream promoter transcription factor (COUP-TF)-interacting protein 2
- HDAC2:
-
Histone deacetylase 2
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Acknowledgments
This work was supported by R01 AI067846 from National Institute of Health/NIAID to Dorina Avram. We thank Adrian Avram for the help with the illustration.
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Albu, D.I., Califano, D., Avram, D. (2010). Flow Cytometry Analysis of Transcription Factors in T Lymphocytes. In: Higgins, P. (eds) Transcription Factors. Methods in Molecular Biology, vol 647. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-738-9_23
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DOI: https://doi.org/10.1007/978-1-60761-738-9_23
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-60761-737-2
Online ISBN: 978-1-60761-738-9
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