Flow cytometry analysis of transcription factors in T lymphocytes.
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.
Albu
Diana I
DI
Center for Cell Biology and Cancer Research, Albany Medical College, Albany, NY, USA.
Califano
Danielle
D
Avram
Dorina
D
D021581:Active Transport, Cell Nucleus; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D015704:CD4 Antigens / Q000378:metabolism; D016827:CD8 Antigens / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D005786:Gene Expression Regulation; D007150:Immunohistochemistry; D008198:Lymph Nodes / Q000166:cytology; D008297:Male; D051379:Mice; D010539:Permeability; D012097:Repressor Proteins / Q000378:metabolism*; D013154:Spleen / Q000166:cytology; D013194:Staining and Labeling; D016176:T-Lymphocyte Subsets / Q000378:metabolism; D013601:T-Lymphocytes / Q000378:metabolism*; D025521:Tumor Suppressor Proteins / Q000378:metabolism*
C511143:Bcl11b protein, mouse; D015415:Biomarkers; D015704:CD4 Antigens; D016827:CD8 Antigens; D012097:Repressor Proteins; D025521:Tumor Suppressor Proteins