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Foxp3 apc

Manufactured by Miltenyi Biotec
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The FoxP3-APC is a fluorescently labeled antibody that binds to the FoxP3 transcription factor, which is a key marker for regulatory T cells. This product can be used for the identification and analysis of regulatory T cells in flow cytometry applications.

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Lab products found in correlation

Cd25 pe, by Miltenyi Biotec (2 mentions) Cd3 efluor 450, by BD (1 mentions) Foxp3 pe, by Miltenyi Biotec (1 mentions) Cd45ra pe, by Miltenyi Biotec (1 mentions) Cd4 fitc, by R&D Systems (1 mentions) Cd4 percp cyanine5, by BD (1 mentions) Cd8 pe cyanine7, by BD (1 mentions) Cd45 apc, by R&D Systems (1 mentions) Cd45ra apc, by Miltenyi Biotec (1 mentions) Cd62l apc, by Miltenyi Biotec (1 mentions) Cd14 pe, by R&D Systems (1 mentions) Cd3 fitc, by BD (1 mentions) Trypan blue staining, by Bio-Rad (1 mentions) Ly6g pe, by Miltenyi Biotec (1 mentions) Cd11b vioblue, by Miltenyi Biotec (1 mentions) Gallios facs analyzer, by Beckman Coulter (1 mentions) Anti cd4 percp, by Miltenyi Biotec (1 mentions) Ly6c fitc, by Miltenyi Biotec (1 mentions) Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions) Foxp3 staining buffer set, by Thermo Fisher Scientific (1 mentions)

4 protocols using foxp3 apc

1

Multicolor Flow Cytometry Analysis of Immune Cells

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Cells were stained with CD3 FITC (Cat# 11-0039), CD3 eFluor 450 (Cat# 48-0038), CD4 PE-Cyanine7 (Cat# 25-0049), CD4 PerCP-Cyanine5.5 (Cat# 560650, BD Pharmingen), Foxp3 APC (Cat# 17-4777), Foxp3 PE (Cat# 12-4776), CD45RA PE (Cat# 130-092-248, Miltenyi), CD45RA APC (Cat# 130-092-249, Miltenyi), CD62L APC (Cat# 17-0629), CD25 PE (Cat# 130-091-024, Miltenyi), CD25 APC-H7 (Cat# 560225, BD Pharmingen), CD8 PE-Cyanine7 (Cat# 25-0088), Perforin PE (Cat# 12-9994), granzyme B PE (Cat# 12-8899), CCR8 APC (Cat# FAB1429A, R&D System), CD4 FITC (Cat# 11-0049), CD45 APC (Cat# 17-9459), CD14 PE (Cat# 12-0149), CD45RO PE (Cat# 12-0457), CD45RO eFluor 450 (Cat# 48-0457), CD127 APC (Cat# 17-1278). PITPNM3 antibody (Cat# NBP1-31070, Novus) was labeled with APC by Abcam APC Conjugation Kit (ab201807, Abcam) according to the manufacturer's instructions. For the intracellular stain, cells were pretreated with Intracellular Fixation and Permeabilization kit (Cat# 88-8824) according to the manufacturer's instructions. All the reagents were from eBioscience unless indicated otherwise. Cells were subsequently analyzed by multicolor flow cytometry (Gallios, Beckman Coulter, China).
Su S., Liao J., Liu J., Huang D., He C., Chen F., Yang L., Wu W., Chen J., Lin L., Zeng Y., Ouyang N., Cui X., Yao H., Su F., Huang J.D., Lieberman J., Liu Q, & Song E. (2017). Blocking the recruitment of naive CD4+ T cells reverses immunosuppression in breast cancer. Cell Research, 27(4), 461-482.
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2

Systemic Inflammation and Leukocyte Trafficking Analysis

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Forty-eight hours or 7 days after CLP procedure, animals were sacrificed by pentobarbital i.p. injection and blood and organs were harvested. Blood count was determined by a hemocytometer, and plasma concentrations of IL1β, IL-6, IL-10, IFNγ, and TNFα were measured by multiplex immunoassays (Bio-Plex Pro Mouse Th1 cytokine, Biorad, France) according to the manufacturer’s recommendations. The same protocol was carried out on healthy mice (H0).
Leukocyte trafficking was analyzed by flow cytometry as previously described [33 (link)]. The spleen and liver were crushed in HBSS and filtered on a 70-μm nylon filter. The bone marrow was extracted from the femur, after the bone has been clipped, by rapidly injecting 1 ml of PBS into the medullary cavity. The lungs were cut into fine pieces and incubated in a cocktail of collagenase I and DNase I at 37 °C for 45 min before being crushed and filtered. After washing, a cell count was performed by a hemocytometer with Trypan blue staining (BioRad). Cell suspensions were labeled with a combination of anti-CD4-PerCP, CD25-PE, CD11b-Vioblue, Ly6C-FITC, Ly6G-PE, FoxP3-APC, and CD45-PerCP mAbs (Miltenyi, France) after permeabilization according to the manufacturer’s recommendations. Data were acquired on a Gallios FACS analyzer (Beckman Coulter). The same protocol was carried out on healthy mice (H0).
Laroye C., Boufenzer A., Jolly L., Cunat L., Alauzet C., Merlin J.L., Yguel C., Bensoussan D., Reppel L, & Gibot S. (2019). Bone marrow vs Wharton’s jelly mesenchymal stem cells in experimental sepsis: a comparative study. Stem Cell Research & Therapy, 10, 192.
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3

Characterization of Regulatory T Cells

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Cells were stained using Foxp3 Staining Buffer Set and the following antibodies from eBioscience: α-CD25-AF-488, α-CTLA-4-PE, α-CD4-PE-Cy5, and α-CD62L-PE-Cy5 or from Miltenyi: Foxp3-APC. Some lymphocytes were labeled with 5 μM carboxyfluorescein succinimidyl ester (CFSE; Invitrogen, Carlsbad, CA) per million cells for 3 min at room temperature. Fluorescent lymph node cells were assessed on a LSRII (BD Biosciences, San Jose, CA, USA) flow cytometer and analyzed with FlowJo software (Tree Star Inc., Ashland, OR, USA). Tregs (CD25hi/Foxp3hi) cells are reported as a percentage of the CD4+ cells with side and forward scatter profiles consistent with live lymphocytes (cells were collected as described earlier; sham n = 4, injury sham = 4).
Rowe R.K., Ellis G.I., Harrison J.L., Bachstetter A.D., Corder G.F., Van Eldik L.J., Taylor B.K., Marti F, & Lifshitz J. (2016). Diffuse traumatic brain injury induces prolonged immune dysregulation and potentiates hyperalgesia following a peripheral immune challenge. Molecular Pain, 12, 1744806916647055.
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4

Immune Cell Profiling in Tumor Samples

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For analyses of immune cell infiltration, the tumors were processed with a tumor dissociation kit (Miltenyi Biotec, Bergisch Gladbach, Germany). Afterwards, the cells were centrifuged with easycoll separating solution (Biochrom) to discard dead cells. The cells were then stained for 30 min at 4 °C with the following fluorescence labeled antibodies: CD4-PCC5.5 (BD Pharmingen), CD8-PE (BD Pharmingen), CD3-V450 (BD Pharmingen), CD11c-PE (BD Pharmingen) and CD45.2-PCC5.5 (eBioscience). Determination of Tregs was performed with FoxP3 Staining Buffer Set and the antibodies CD4-Vioblue, CD25-AF488 and FoxP3-APC (Miltenyi Biotec). Multicolor flow cytometry was performed with the Gallios Flow Cytometer (Beckman Coulter Inc.).
Werthmöller N., Frey B., Wunderlich R., Fietkau R, & Gaipl U.S. (2015). Modulation of radiochemoimmunotherapy-induced B16 melanoma cell death by the pan-caspase inhibitor zVAD-fmk induces anti-tumor immunity in a HMGB1-, nucleotide- and T-cell-dependent manner. Cell Death & Disease, 6(5), e1761-.
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