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Cd8 fitc

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CD8-FITC is a fluorescently labeled monoclonal antibody that binds to the CD8 surface antigen expressed on a subset of T lymphocytes. It is commonly used in flow cytometry applications for the identification and enumeration of CD8-positive cells.

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Lab products found in correlation

Cd4 pe, by BD (16 mentions) Cd4 percp, by BD (11 mentions) Cd3 percp, by BD (11 mentions) Cd3 apc, by BD (10 mentions) Cd3 pe, by BD (10 mentions) Facscalibur, by BD (10 mentions) Cd25 pe cy7, by BD (8 mentions) Cd4 apc, by BD (8 mentions) Pd 1 pe, by BD (7 mentions) Cd4 fitc, by BD (7 mentions) Cd4 apc cy7, by BD (7 mentions) Cd45ro apc, by BD (6 mentions) Ifn γ pe, by BD (6 mentions) Facscalibur flow cytometer, by BD (6 mentions) Cd27 fitc, by BD (6 mentions) Cd19 pe cy7, by BD (6 mentions) Lsrfortessa, by BD (6 mentions) Cd4 apc h7, by BD (5 mentions) Cd56 apc, by BD (5 mentions) Cd3 v450, by BD (5 mentions)

Close spelling variants of this product

FITC-conjugated anti-CD8 (15 citations) FITC anti-mouse CD8 (8 citations) FITC-conjugated anti-human CD8 mAb (7 citations) FITC Mouse Anti-Human CD8 (6 citations) Anti-human CD8-FITC (6 citations) FITC-labeled anti-human CD8 (3 citations) FITC-conjugated anti-human CD8 antibody (3 citations) CD3 FITC/CD8 PE/CD45 PerCP/CD4 APC (3 citations) FITC-rat anti-mouse CD8 (3 citations) CD8-APC/FITC (3 citations)

132 protocols using cd8 fitc

1

Cross-presentation Experiments with Antibodies

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The following antibodies were used in the cross-presentation experiments: TruStain Fcblock anti-mouse CD16/32 (101320; BioLegend), FITC-CD11c (117306; Biolegend), APC-H2Kb-bound SIINFEKL (141606; Biolegend), APC/Cy7-CD40 (124637; Biolegend), PerCP/Cy5.5- ICAM_1 (116123; Biolegend), BV510-CD86 (563077; BD), PE/Cy7-MHC-II (107629; Biolegend), V450-CD80 (12519; BD). The following antibodies were used for the immunological analysis in the in vivo animal experiments: FITC-CD8 (553062; BD), PE-CD4 (100408; Biolegend), APC-CXCR3 (562266; BD), PerCP/Cy5.5-CD3 (100732; Biolegend), PE-CXCR3 (155903); FITC-CD8 (11083782); PerCP/Cy5.5-CD3 (100328);. Flow cytometric analyses were performed using Fortessa LSR Flow Cytometer (BD Biosciences) or BD Accuri C6 Plus (BD Bioscience). FlowJo software v.10 (FlowJo) was used for the data analysis.
Feola S., Russo S., Martins B., Lopes A., Vandermeulen G., Fluhler V., De Giorgi C., Fusciello M., Pesonen S., Ylösmäki E., Antignani G., Chiaro J., Hamdan F., Feodoroff M., Grönholm M, & Cerullo V. (2022). Peptides-Coated Oncolytic Vaccines for Cancer Personalized Medicine. Frontiers in Immunology, 13, 826164.
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2

Isolating CD4+ and CD8+ T Cells

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CD4+ T cells were isolated by negative selection from freshly obtained PBMC using human CD4+ T cell isolation kit (Miltenyi Biotech, Bergisch Gladbach, Germany). Purity was confirmed with CD4-FITC (Cat# 555346, BD Bioscience) and analyzed by flow cytometry. CD8+ T cells were isolated by negative selection from freshly obtained PBMC using human CD8+ T cell isolation kit (Miltenyi Biotech, Bergisch Gladbach, Germany). Purity was confirmed with CD8-FITC (Cat# 555366, BD Bioscience) and analyzed by flow cytometry. Cells were cultured in RPMI 1640 medium (Sigma) supplemented with 10% heat-inactivated endotoxin-tested FCS (Biochrom GmbH, Berlin, Germany).
Hart M., Walch-Rückheim B., Friedmann K.S., Rheinheimer S., Tänzer T., Glombitza B., Sester M., Lenhof H.P., Hoth M., Schwarz E.C., Keller A, & Meese E. (2019). miR-34a: a new player in the regulation of T cell function by modulation of NF-κB signaling. Cell Death & Disease, 10(2), 46.
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3

Quantifying T-cell Subsets in Blood

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In order to quantify T lymphocyte subsets, 100 μl of whole blood was incubated in 900 μl of Tris-NH4Cl buffer (Thermo Fisher Scientific) at room temperature for 5 min to lyse erythrocytes. After two washes with phosphate-buffered saline (PBS), cells were incubated with CD3-APC, CD4-PerCP, and CD8-FITC antibodies (5 μg/ml each, all from BD Biosciences) for 15 min on ice. After another two washes with PBS, cells were resuspended in 500 μl of PBS. Samples were analyzed on a BD FACS Canto Plus flow cytometer. Among all collected events, single events were gated between forward scatter (FSC)-A and FSC-H. Cell debris was excluded, and intact cells were then gated from single events based on FSC-A and side scatter (SSC). Each cell population was then detected based on antibody staining.
Qun S., Wang Y., Chen J., Huang X., Guo H., Lu Z., Wang J., Zheng C., Ma Y., Zhu Y., Xia D., Wang Y., He H., Wang Y., Fei M., Yin Y., Zheng M., Xu Y., Ge W., Hu F, & Zhou J. (2020). Neutrophil-to-Lymphocyte Ratios Are Closely Associated With the Severity and Course of Non-mild COVID-19. Frontiers in Immunology, 11, 2160.
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4

Multiparametric Flow Cytometry Analysis

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Pooled splenocytes (2×105 cells/group, n = 3-4 / group) obtained from naive control, MPTP and calpeptin treated mice were stained with CD8-FITC (BD Biosciences, San Jose, CA) and CD4-PerCP (BD Biosciences, San Jose, CA) as described [20 (link)]. Flow cytometric two-parameter dot plots and quadrant statistics were generated using FACScan and CellQuest software (BD Biosciences, Mountain view, CA). For intracellular staining, cells were fixed and permeabilized using Fix and Perm reagents (BD Biosciences, San Jose, CA), and then incubated with specific antibodies [75 (link)]. Briefly, 5×105 cells per group were re-suspended in 0.5 mL of Fixation/Permeabilization Buffer (BD) and incubated at 2-8° C for 30 minutes. After washing, the cell pellet was re-suspended in Permeabilization/Wash Buffer and stained with Foxp3-APC (BD Biosciences, San Jose, CA). Cells were then analyzed on FACScan using CellQuest software as described above.
Samantaray S., Knaryan V.H., Shields D.C., Cox A.A., Haque A, & Banik N.L. (2015). Inhibition of calpain activation protects MPTP-induced nigral and spinal cord neurodegeneration, reduces inflammation, and improves gait dynamics in mice. Molecular neurobiology, 52(2), 1054-1066.
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5

NY-ESO-1-specific T Cell Activation in Alginate Cryogels

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CD8+ T cells from the
same HLA-A2.1+ donor as the moDCs were transfected, as
previously described27 (link) with mRNA (5 mg/mL,
BioNTech RNA Pharmaceuticals) encoding for a mouse T cell receptor
(specific for the HLA-A2.1-specific NY-ESO-1 epitope SLLWITQC). Transfection
efficiency was determined 1 day after transfection using anti-mouse
TCR-β-FITC (BioLegend) and was typically between 80 and 90%
(Figure S9A). These NY-ESO-1 specific T
cells were stained with CellTrace Violet and were added to activated
moDCs (ratio DCs/T cells as 1:2.5) in alginate cryogels. The supernatant
was taken after 24 h of T cell activation to determine IFNγ
production. After 72 h of activation, cells were collected and stained
with Fixable Viability Dye eFluor 780, CD8-FITC (BD BioScience), and
CD25-PE (BD BioScience).
Weiden J., Schluck M., Ioannidis M., van Dinther E.A., Rezaeeyazdi M., Omar F., Steuten J., Voerman D., Tel J., Diken M., Bencherif S.A., Figdor C.G, & Verdoes M. (2021). Robust Antigen-Specific T Cell Activation within Injectable 3D Synthetic Nanovaccine Depots. ACS Biomaterials Science & Engineering, 7(12), 5622-5632.
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6

PBMC Isolation and Cell Sorting

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Peripheral blood samples were collected at baseline and at weeks 12 and 24 of treatment. PBMCs were isolated by density-gradient centrifugation according to standard protocols and were cryopreserved in liquid nitrogen until analysis. For cell sorting, the PBMCs (0.1–1 × 107) were thawed, washed with RPMI-1640 containing 10–20% FBS, and then treated with 100 U/mL DNase I (Roche) to degrade DNA released from dead or dying cells in order to reduce cell clumps. The remaining PBMCs were labeled with Live/Dead (Life Technologies), CD4-APC, CD8-FITC, and CD19-PE (BD Biosciences) and were sorted using a FACSAria III Cell Sorter (BD Biosciences). The staining profile and gating strategy for cell sorting are shown in Figure S1 in Supplementary Material.
Xu Y., Liu Y., Zhao M., Chen Y., Xie C., Gong M., Deng H., Li X., Sun J., Hou J., Wu H, & Wang Z. (2017). Dynamic Perturbations of CD4 and CD8 T Cell Receptor Repertoires in Chronic Hepatitis B Patients upon Oral Antiviral Therapy. Frontiers in Immunology, 8, 1142.
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7

Comprehensive Immune Cell Profiling

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The monoclonal antibodies used in this study included CD3 PerCP, CD5 APC-R700, CD8-FITC, CD11c PE-Cy5, CD14 APC-H7, CD19 PE-Cy5, CD25 PE-Cy7, CD24 FITC, CD27 PE-Cy7, CD28 PE-Cy5, CD38 APC-H7, CD45 V500, CD45RO APC, CD56 APC, CD57 FITC, CD80 PE, CD86 APC, CD123 APC, CD127 BV421, CCR7 PE, TIGIT BV421, HLA DR V450, TIM3 PE, CXCR5 APC-R700, PD1 BV421, and PDL1 PE-Cy7 (BD Biosciences).
Rodero-Romero A., Sainz de la Maza S., Fernández-Velasco J.I., Monreal E., Walo-Delgado P.E., Chico-García J.L., Villarrubia N., Rodríguez-Jorge F., Rodríguez-Ramos R., Masjuan J., Costa-Frossard L, & Villar L.M. (2023). Blood CD8+ Naïve T-Cells Identify MS Patients with High Probability of Optimal Cellular Response to SARS-CoV-2 Vaccine. Vaccines, 11(9), 1399.
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8

Comprehensive T-cell Signaling Antibody Panel

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Antibodies and reagents used for flow cytometry were: CD8-APC, Fixable viability stain 450, antiCD16/32 (eBiosciences); anti-rabbit-PE (Jackson Immunoresearch); anti-SLP76 (pY128), CD4-PE, CD25-FITC, CD8 FITC and anti-mouse IgG1-PE (BD Biosciences); CD4-PEVio770 (Miltenyi Biotec); CD3-BV421 (Biolegend); anti-pErk1/2 (pT202/pY204) (Cell signaling Technology). The following reagents were used for cell stimulation, immunoblotting or immunoprecipitation: anti-CD3-biontinylated, anti-CD28-biontinylated, anti-CD3ε, anti-CD28 (eBiosciences); streptavidin (Sigma); anti-pErk1/2 (pT202/pY204), anti-pPLCγ1 (pY783), anti-pp38 (pT180/pY182), anti-pJNK (pT183/Y185), anti-pAkt (pS473), anti-pSLP76 (pS376) (Cell Signaling Technology); anti-GADS (Santa Cruz Biotechnology Inc.); rabbit anti-SLP76 or goat-anti-SLP76 (Thermo Fisher Scientific); anti-β-tubulin (Chemicon); for anti-14-3-3 immunoblotting we used a mix of anti-pan 14-3-3 and anti-14-3-3ζ (Santa Cruz, Biotechnology Inc.).
Navas V.H., Cuche C., Alcover A, & Di Bartolo V. (2017). Serine Phosphorylation of SLP76 Is Dispensable for T Cell Development but Modulates Helper T Cell Function. PLoS ONE, 12(1), e0170396.
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9

T-Cell Activation and Monocyte Stimulation Assay

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Both PMNs and CD14+ monocytes were stimulated for 10 min with 4 μM calcium ionophore A23178 (Merck, Darmstadt, Germany). CD4+ cells were activated using αCD3/CD28 beads (Gibco). Purity and activation were checked by FACS (LSRIII, BD, San Jose, USA) staining the cells with CD3-PE (clone SK7)/CD4-APC (clone SK3)/CD8-FITC (clone SK1)/CD14-PEcy7 (clone M5E2) and activation using CD25-AF700 (clone BC96). The percentage of CD25+ cells was > 35%.
Alarcon-Barrera J.C., von Hegedus J.H., Brouwers H., Steenvoorden E., Ioan-Facsinay A., Mayboroda O.A., Ondo-Mendez A, & Giera M. (2020). Lipid metabolism of leukocytes in the unstimulated and activated states. Analytical and Bioanalytical Chemistry, 412(10), 2353-2363.
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10

Comprehensive Immune Profiling Workflow

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ELISA kits for murine IL-6 was from R&D Systems (Minneapolis, MN). Fluorescein-conjugated mAbs including anti-CD3-PE-Cy7, CD4-PE-Cy5, CD8-PE, CD8-FITC, CD11b-APC, Ly6G-PE, Ly6C-FITC, CXCR2-Percp-Cy5.5, CD45-BV510, PD1-PE, PDL1-PE, LAG3-PE, CTLA4-PE, IFNγ-PE and isotype antibodies were purchased from BD biosciences. TIM3-PE was from Miltenyi Biotec (Bergisch-Gladbach, Germany). Antibody against STAT3 (79D7, 4904S), p-STAT3 (Tyr705, 9131S), ZEB1 (3396P), ZO-1(5406P), Snail(3879P), N-Cadherin(4061P) were obtained from Cell Signaling Technology (Beverly, MA, USA) and antibody against Actin was from Santa Cruz. IL6 inhibitor (501109) and IFN-γ inhibitor (505827) was from Biolegend. Luciferin substrate (K9909PE) for in vivo image was purchased from PerkinElmer Inc (Hopkinton, MA, USA). HE staining kit was from Beyotime Biotechnology. ShRNA for ZEB1 (TG513177) was purchased from OriGene.
Zhu H., Gu Y., Xue Y., Yuan M., Cao X, & Liu Q. (2017). CXCR2+ MDSCs promote breast cancer progression by inducing EMT and activated T cell exhaustion. Oncotarget, 8(70), 114554-114567.
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