How do you stain FoxP3?
Wash 2x with freshly made, cold 1x Permeabilization/Wash Buffer. Add FoxP3 antibody (or intracellular cytokines) to cells and incubate for 30 minutes at 4 °C. Wash cells 1x with cold 1x Permeabilization/Wash Buffer. Resuspend cells in Flow Cytometry Staining Buffer and analyze on a flow cytometer.
Is FoxP3 intracellular?
88-8996-40). Cells in the lymphocyte gate were used for analysis. Foxp3 protein is difficult to stain since it is not very abundant, and it is an intracellular protein.
What is permeabilization buffer?
Description. The RayBiotech Cell Permeabilization Buffer is saponin-containing buffer commonly used to permeabilize cells and allow antibody to stain target intracellularly.
What cells express FOXP3?
Forkhead box P3 (Foxp3) is an important transcription factor that belongs to the forkhead/winged-helix family of transcriptional regulators. However, recent work has proposed that Foxp3 can be expressed by other types of lymphoid cells or myeloid cells and also by some non-hematopoietic cells such as epithelial cells.
What are FOXP3 Tregs?
FOXP3+ regulatory T (Treg) cells are critical in maintaining immune tolerance and homeostasis of the immune system. The molecular mechanisms underlying the stability, plasticity and functional activity of Treg cells have been much studied in recent years.
How do you prepare a permeabilization buffer?
A. 1X Phosphate Buffered Saline (PBS): To prepare 1 L 1X PBS: add 100 ml 10X PBS (#12528) to 900 ml water, mix. Cell Permeabilization Buffer: Purchase ready-to-use (#39487) or to prepare 10 ml, add 30 µl Triton™ X-100 to 10 ml Antibody Dilution Buffer. Store at 4°C.
What is FC block?
Mouse BD Fc Block is a purified rat IgG 2b anti-mouse CD16/CD32 monoclonal antibody (Cat. 550270 and 550271) is a purified mouse IgG 1anti-rat CD32 monoclonal antibody. 2 Mouse and Rat BD Fc Block can be used to block the Fc-mediated adherence of antibodies* to mouse and rat FcR, respectively.
What is FBS buffer?
Description. Stain Buffer (FBS) can be used for the immunofluorescent staining of single-cell suspensions prepared from either lymphoid tissues, bone marrow, peripheral blood, or cultured cells. In addition, Stain Buffer (FBS) contains the metabolic inhibitor, sodium azide (NaN3).
What regulates FOXP3?
FOXP3 has been shown to interact with the acetyltransferase Tat-interaction protein 60 kDa (TIP60) to promote FOXP3 acetylation, which is required for Treg cells’ suppressive function (19). P300 also regulates FOXP3 acetylation and stability by preventing proteasome-mediated FOXP3 degradation (20).
What does FOXP3 code for?
The FOXP3 gene provides instructions for producing the forkhead box P3 (FOXP3) protein. The FOXP3 protein attaches (binds) to specific regions of DNA and helps control the activity of genes that are involved in regulating the immune system.