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Cd25 pe cf594

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CD25-PE-CF594 is a fluorescently-labeled antibody that binds to the CD25 cell surface antigen. It can be used for the identification and enumeration of CD25-positive cells in flow cytometry applications.

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Lab products found in correlation

Cd4 buv395, by BD (2 mentions) Horizon brilliant stain buffer, by BD (1 mentions) Cd45 apc h7, by BD (1 mentions) Cd4 bv510, by BD (1 mentions) Falcon round bottom polystyrene tubes, by Corning (1 mentions) Cd3 percp clone sk7, by BD (1 mentions) Cd8 pe clone hit8a, by BD (1 mentions) Cd14 bv605, by BD (1 mentions) Facs lysing solution, by BD (1 mentions) Facsymphony a5 cytometer, by BD (1 mentions) Cd45ra pe cy7, by BD (1 mentions) Cd127 apc, by BioLegend (1 mentions) Cd8 bv785, by BD (1 mentions) Foxp3 pe, by Thermo Fisher Scientific (1 mentions) Live dead ghost dye 780, by Cytek Biosciences (1 mentions) Cd62l bv605, by BD (1 mentions) Cd44 pe cy7, by BD (1 mentions) Cd3 fitc, by BD (1 mentions) Cd45 bv510, by BD (1 mentions) Anti mouse cd16 cd32 antibody, by BD (1 mentions)

Close spelling variants of this product

PE-CF594 anti-CD25 PE-CF594 CD25 (M-A251; Anti-CD25-PE-CF594 (clone M-A251) CD25-PE

5 protocols using cd25 pe cf594

1

Comprehensive Immunophenotyping of Peripheral Blood

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PB samples were collected through venipuncture in 2 mL BD Vacutainer® spray-coated K2EDTA blood collection tubes (BD Biosciences, San Jose, CA, USA; #367841). One hundred μL of whole blood was transported into 5 mL Corning™ Falcon™ Round-Bottom Polystyrene Tubes (#352054) and stained with 12 monoclonal antibodies against the surface markers: CD45-APC-H7 (clone 2D1, #560178); CD3-PerCP (clone SK7, #340663); CD4-BV510 (clone SK3; #562970); CD8-PE (clone HIT8a, #555635); CD14-BV605 (clone M5E2, #564054); CD16-APC (clone B73.1, #561304); CD56-APC-R700 (clone NCAM16.2, #565139); CD25-PE-CF594 (clone M-A25, #562403); CD11b-BV786 (clone D12, #742642); CD183 (CXCR3)-BV421 (clone 1C6/CXCR3, #562558); CD194 (CCR4)-BV650 (clone 1G1, #744140); and CD196 (CCR6)-BB515 (clone 11A9, #564479) (all from BD Biosciences). The staining procedure was performed in BD Horizon™ Brilliant Stain Buffer (BD Biosciences, #563794) followed by red blood cells lysis with 1x BD FACS™ lysing solution (BD Biosciences, #349202) as suggested by the manufacturer for the Lyse-no-Wash protocol.
Orologas-Stavrou N., Politou M., Rousakis P., Kostopoulos I.V., Ntanasis-Stathopoulos I., Jahaj E., Tsiligkeridou E., Gavriatopoulou M., Kastritis E., Kotanidou A., Dimopoulos M.A., Tsitsilonis O.E, & Terpos E. (2020). Peripheral Blood Immune Profiling of Convalescent Plasma Donors Reveals Alterations in Specific Immune Subpopulations Even at 2 Months Post SARS-CoV-2 Infection. Viruses, 13(1), 26.
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2

Comprehensive Immunophenotyping of CLL Samples

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PBMCs from each CLL donor were fixed, barcoded,34 and stained for 30 min with the antibody panel. The following antibodies were used: CD3-BUV395, CD4-BUV563, HLA-DR-BUV615, CD16-BUV737, CXCR5 (CD185)-BUV805, CCR7 (CD197)-BV605, CCR6 (CD196)-BV711, CD56-BV750, CD127-BV786, PD-1 (CD279)-BB700, CD14-PE, CD25-PE-CF594, CCR3 (CD183)-PE-Cy5, CD45RA-PE-Cy7, CD69-APC-R700, CD8/CD19-APC-Cy7 (BD Biosciences). Experiments were analyzed with a BD FACSymphony A5 cytometer (BD Biosciences) and further processed in Cytobank (https://cellmass.cytobank.org/cytobank/). The FlowSOM clustering algorithm was applied to identify cell populations, which were validated by manual gating. The UMAP dimensionality reduction algorithm was used to visualize the data.
Yin Y., Athanasiadis P., Karlsen L., Urban A., Xu H., Murali I., Fernandes S.M., Arribas A.J., Hilli A.K., Taskén K., Bertoni F., Mato A.R., Normant E., Brown J.R., Tjønnfjord G.E., Aittokallio T, & Skånland S.S. (2022). Functional testing to characterize and stratify PI3K inhibitor responses in chronic lymphocytic leukemia. Clinical cancer research : an official journal of the American Association for Cancer Research, 28(20), 4444-4455.
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3

Immunotherapy for Prostate Cancer in Mice

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Six-week-old FVB/NJ mice were implanted with 1×106 MyC-CaP cells administered subcutaneously. Only male mice were used for these studies, given that the tumor of origin is male specific. Each mouse was then immunized intradermally with 100 μg DNA vaccine (pTVG-AR), or vector control (pTVG-4) weekly, beginning 1 day after tumor implantation. TLR agonists were coadministered with the vaccine intradermally at the following doses: TLR3 (Poly(I:C) HMW, 100 µg/mouse), TLR9 (ODN 1826, 50 µg/mouse). Mice were further treated with immune checkpoint blockade, αPD-1, αLAG-3, αCTLA-4, or IgG control the day following each immunization. Tumor volumes were measured as described above. In a parallel study, tumors were collected after treatment and digested in collagenase, DNAse I, and protease inhibitors for 2 hours at 37°C, passed through a 100 mm mesh screen, and analyzed by flow cytometry with the following antibodies: CD3-FITC (BD 555274), CD4-BUV395 (BD 563790), CD8-BV785 (BD 563332), CD25-PE-CF594 (BD 562694), CD44-PE-Cy7 (BD 561283), CD45-BV510 (BD 563891), CD62L-BV605 (BD 563252), CD127-APC (Biolegend 121122), KLRG1-BV711 (BD 564014), FOXP3-PE (Thermo Fisher 12-5773-82), 4-1BB-PerCP-eF710 (Lifetech 46-1371-82), and Live/Dead Ghost dye 780 (Tonbo, San Diego, CA 13-0865-T100).
Jeon D., Hill E., Moseman J.E, & McNeel D.G. (2024). Combining toll-like receptor agonists with immune checkpoint blockade affects antitumor vaccine efficacy. Journal for Immunotherapy of Cancer, 12(5), e008799.
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4

Isolation and Flow Cytometry Analysis of Leukocytes

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Leukocytes from lymph nodes, blood, spleen and peritoneal cavity wash were collected and isolated as described [56 (link)], and analysed by flow cytometry. Anti-mouse CD16/CD32 antibody (1:50, BD Biosciences, clone #2.4G2) was used to block non-specific binding. The following fluorochrome-conjugated antibodies were used: CD3e-AF700 (1:200, BD Biosciences, clone #500A2), CD4-BUV395 (1:100, BD Biosciences, clone #RM4-5), CD8a-PerCP (1:100, BioLegend, clone #53-6.7), CD25-PE-CF594 (1:100, BD Biosciences, #PC-61), FoxP3-APC (1:50, eBioscience, clone #FJK-16S), CD11b-APC (1:200, BioLegend, clone #MI/71), CD11c-BV421 (1:50, BD Bioscience, clone #HL3), GR1-PerCP Cy5.5 (1:200, BD Biosciences, clone #RB6-8C5), F4/80-BV711 (1:100, BioLegend, clone #BM8) and MHCII-APC/Cy7 (1:500, BioLegend, clone #M5/114.15.2). Single-stained beads were used to set compensation controls, and fluorescence-minus-one (FMO) controls were used to define population gates. Data were acquired using a BD LSRFortessa X-20 (BD Biosciences), and were analysed using FlowJo software v10.5.0 (BD Biosciences).
Wilson A.L., Wilson K.L., Bilandzic M., Moffitt L.R., Makanji M., Gorrell M.D., Oehler M.K., Rainczuk A., Stephens A.N, & Plebanski M. (2018). Non-Invasive Fluorescent Monitoring of Ovarian Cancer in an Immunocompetent Mouse Model. Cancers, 11(1), 32.
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5

Phenotyping Human PBMC Subsets

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Single-cell suspensions from human PBMCs, prepared as below, were washed in flow cytometry staining buffer (PBS with 2% FCS and 0.02% sodium azide) followed by blocking with 10% FCS. The following mAbs from BD Biosciences were used at optimally titrated concentrations: CD1d-allophycocyanin (51.1), CD5-V450 (UCHT2), CD19-FITC (HIB19), CD24-PerCP-Cy5.5 (ML5), CD25-PE-CF594 (M-A251), CD27-V500 (M-T271), CD38-AF700 (HIT2). Viable cells were distinguished using Fixable Viability Dye eFluor R 780 (eBioscience).
Populations of interest were gated as above. Representative gating strategy is illustrated in Supporting Information Figure 2. Samples were acquired on an LSR Fortessa flow cytometer (BD Biosciences) and were analyzed with FlowJo software (Tree Star).
Khan A.R., Amu S., Saunders S.P., Hams E., Blackshields G., Leonard M.O., Weaver C.T., Sparwasser T., Sheils O, & Fallon P.G. (2015). Ligation of TLR7 on CD19(+) CD1d(hi) B cells suppresses allergic lung inflammation via regulatory T cells. European journal of immunology, 45(6).
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