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Treg permeabilization buffers

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Treg permeabilization buffers are a set of solutions designed to facilitate the intracellular staining of regulatory T cells (Tregs) for flow cytometric analysis. These buffers allow for the permeabilization of cell membranes, enabling the detection of intracellular Treg-specific markers such as Foxp3.

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Lab products found in correlation

Rat igg2b eb149 10h5, by Thermo Fisher Scientific (2 mentions) Cytofix cytoperm kit, by BD (2 mentions) Cd11c, by Thermo Fisher Scientific (1 mentions) Foxp3 fjk 16s, by Thermo Fisher Scientific (1 mentions) Live dead aqua, by Thermo Fisher Scientific (1 mentions) Thy 1, by Thermo Fisher Scientific (1 mentions) Cd62l, by Thermo Fisher Scientific (1 mentions) Cd11b, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Annexin 5, by Thermo Fisher Scientific (1 mentions) Clip 15g4, by Santa Cruz Biotechnology (1 mentions) Rmp1 30, by Thermo Fisher Scientific (1 mentions) Ebioy ae, by Thermo Fisher Scientific (1 mentions)

2 protocols using treg permeabilization buffers

1

Comprehensive Immune Cell Phenotyping

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Antibodies used include: Ii(In-1, 1:100), H2-M (2E5A, 1:50), Rat IgG1 (R3–34, 1:50) (all from BD Biosciences); podoplanin (8.1.1, Biolegend, 1:500); CLIP (15G4, Santa Cruz, 1:5); H2-O69 (link) (Mags.Ob3, Lisa Denzin, 1:400); 10.1.1 (UVA lymphocyte culture center, 1:1000); CD45 (30-F11, 1:1000), CD31 (390, 1:1000), MHC-II (M5/114.15.2, 1:50 (Fig 1) to 1:500 (Fig 6)), CD8 (53-6.7, 1:1000), CD4 (GK1.5, 1:1000), Thy1.1 (HIS51, 1:1000), Thy1.2 (53-2.1, 1:1000), CD45.1 (A20, 1:500), CD11c (N418, 1:500), CD11b (M1/70, 1:500), CD69 (H1.2F3, 1:500), CD62L (MEL-14, 1:1000), CD44 (IM7, 1:1000), CD25 (PC61.5, 1:500), Y-AE (eBioY-Ae, 1:200), PD-1 (RMP1–30, 1:100), LAG-3 (eBioC9B7W, 1:100), Rat IgG2b (eB149/10H5) (all from eBioscience). Intracellular staining for PD-1, LAG-3, Ii, H2-M and H2-O was done using the Cytofix/Cytoperm kit (BD Bioscience), and Ki67 (SolA15, 1:500) and FoxP3 (FJK-16s, 1:100) were stained using Treg permeabilization buffers (eBioscience). Annexin V (1:20) was stained using the eBioscience kit. DAPI (Sigma) or live/dead aqua (Invitrogen, 1:200) were used to distinguish live cells. Cells were acquired on a FACSCanto II (BD Biosciences) and data analyzed using FlowJo (Tree Star).
Rouhani S.J., Eccles J.D., Riccardi P., Peske J.D., Tewalt E.F., Cohen J.N., Liblau R., Mäkinen T, & Engelhard V.H. (2015). Roles of lymphatic endothelial cells expressing peripheral tissue antigens in CD4 T-cell tolerance induction. Nature Communications, 6, 6771.
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2

Multicolor Flow Cytometry Panel

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Antibodies used include: Ii (In-1, 1:100), H2-M (2E5A, 1:50), Rat IgG1 (R3-34, 1:50; all from BD Biosciences); podoplanin (8.1.1, BioLegend, 1:500); CLIP (15G4, Santa Cruz, 1:5); H2-O (ref. 69 (link); Mags.Ob3, Lisa Denzin, 1:400); 10.1.1 (UVA lymphocyte culture centre, 1:1,000); CD45 (30-F11, 1:1,000), CD31 (390, 1:1,000), MHC-II (M5/114.15.2, 1:50 (Fig. 1) to 1:500 (Fig. 6)), CD8 (53-6.7, 1:1,000), CD4 (GK1.5, 1:1,000), Thy1.1 (HIS51, 1:1,000), Thy1.2 (53-2.1, 1:1,000), CD45.1 (A20, 1:500), CD11c (N418, 1:500), CD11b (M1/70, 1:500), CD69 (H1.2F3, 1:500), CD62L (MEL-14, 1:1,000), CD44 (IM7, 1:1,000), CD25 (PC61.5, 1:500), Y-AE (eBioY-Ae, 1:200), PD-1 (RMP1-30, 1:100), LAG-3 (eBioC9B7W, 1:100), Rat IgG2b (eB149/10H5; all from eBioscience). Intracellular staining for PD-1, LAG-3, Ii, H2-M and H2-O was done using the Cytofix/Cytoperm kit (BD Bioscience), and Ki67 (SolA15, 1:500) and FoxP3 (FJK-16s, 1:100) were stained using Treg permeabilization buffers (eBioscience). Annexin V (1:20) was stained using the eBioscience kit. 4,6-Diamidino-2-phenylindole (DAPI; Sigma) or live/dead aqua (Invitrogen, 1:200) were used to distinguish live cells. Cells were acquired on a FACSCanto II (BD Biosciences) and data analysed using FlowJo (Tree Star).
Rouhani S.J., Eccles J.D., Riccardi P., Peske J.D., Tewalt E.F., Cohen J.N., Liblau R., Mäkinen T, & Engelhard V.H. (2015). Roles of lymphatic endothelial cells expressing peripheral tissue antigens in CD4 T-cell tolerance induction. Nature Communications, 6, 6771.
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