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Cd8α apc cy7

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CD8α-APC-Cy7 is a fluorochrome-conjugated antibody that binds to the CD8α cell surface marker. It is designed for use in flow cytometry applications to identify and quantify CD8-positive cells within a sample.

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Lab products found in correlation

Cd11c apc, by BD (2 mentions) Cd4 v500, by BD (1 mentions) Mhc 2 pe, by BD (1 mentions) Mhc 1 fitc, by BD (1 mentions) Ifn γ apc cy7, by BD (1 mentions) Cd86 pe cy7, by BD (1 mentions) Cd3 fitc, by BD (1 mentions) Cd44 apc, by BD (1 mentions) Cd11b v500, by BD (1 mentions) Fcr blocking reagent, by Miltenyi Biotec (1 mentions) B220 percp, by BD (1 mentions) Cd4 alexa700, by BD (1 mentions) αβtcr pe, by BD (1 mentions) Cd45 pe cy7, by BD (1 mentions) Moflo sorter, by Beckman Coulter (1 mentions) Cd11b pe, by BD (1 mentions) E cadherin pe, by BioLegend (1 mentions) Cd11b percp cy5, by BD (1 mentions) Aria sorp, by BD (1 mentions) Fortessa, by BD (1 mentions)

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APC-Cy7 (76 citations) APC-CD8α (4 citations)

4 protocols using cd8α apc cy7

1

Multiparameter Flow Cytometry of Immune Cells

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Prior to flow cytometry, cells were washed in staining buffer (0.05% (w/v) BSA, 2 mM EDTA in 1× PBS) and treated with FcR blocking reagent (Miltenyi Biotec) for 10 min. Subsequently, In vitro differentiated single cell clones and Dox-pDC were stained with the following antibodies for 30 min at 4°C: CD11c-APC, MHC-I-FITC, MHC-II-PE, SiglecH-PE, CD86-PE-Cy7, CD289 (TLR9)-FITC, CD11b-V500, B220-PerCP, CD8α-APC-Cy7 (all BD Biosciences) and CD9-FITC (Thermo Fisher). T lymphocytes were stained with the following antibodies: CD3-FITC, CD4-V500, CD8α-APC-Cy7, CD44-APC and IFNγ-APC-Cy7 (all BD Biosciences); CD62L-PerCP-Cy5.5 and RORγt-PerCP-ef710 (all Thermo Fisher Scientific). Flow cytometry was performed using LSRII and FlowJo analysis software (V10; FlowJo, Ashland, USA).
Thieme S., Holzbaur A., Wiedemuth R., Binner A., Navratiel K., Anastassiadis K., Brenner S, & Richter C. (2018). The Dox-pDC - A murine conditionally immortalized plasmacytoid dendritic cell line with native immune profile. PLoS ONE, 13(2), e0192437.
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2

Engraftment Evaluation of Human HSPCs

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Human HSPC engraftment in NSG recipients was evaluated by peripheral blood (PB) lymphocyte typing using seven-color LSR II SORP flow cytometry 30 days after transplantation. Briefly, whole blood from NSG recipients was collected in heparinized tubes, and aliquots of 100 μl were stained with human-specific CD45 PE-Cy7, along with a combination of the following human mAb (BD Biosciences): CD8α APC-Cy7, CD4 Alexa 700, αβTCR PE, γδTCR PE, CD56 APC, CD19 APC, CD11c Alexa 700, HLA-DR PerCP and CD33 FITC. Tissue typing of PB lymphocyte, spleen and BM was performed 180 days after transplantation.
Huang Y., Elliott M.J., Yolcu E.S., Miller T.O., Ratajczak J., Bozulic L.D., Wen Y., Xu H., Ratajczak M.Z, & Ildstad S.T. (2015). Characterization of human CD8+TCR- facilitating cells in vitro and in vivo in a NOD/SCID/IL2rγnull mouse model. American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons, 16(2), 440-453.
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3

Isolation and Sorting of Immune Cells

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Splenocytes were prepared from 12-week mice, erythrocytes were removed by incubation in lysis buffer (1:9 v/v 0.17M Tris: 0.16 M ammonium chloride) for 5 min and enriched splenocytes were resuspended in 2% FBS and 1 mM EDTA in Hank’s Balanced Salt Solution. For isolation of tumour lineages, dissected gp130F/F gastric tumours were chopped into ~3–4mm3 pieces and disaggregated non-enzymatically by incubation in dissociation buffer (5% FBS, 1 mM dithiothreitol, 1 mM EDTA in PBS) for 1 h at 37 °C with agitation. Digested tissue pieces were passed through a 70-μm strainer, and the cells were resuspended in 2% FBS and 2 mM EDTA in Hank’s Balanced Salt Solution. Splenocytes and gastric tumour cells were stained with CD11c-APC (1:500), CD8α-APC-Cy7 (1:500), CD45R/B221-FITC (1:500), CD11b-PE (1:500), Gr-1 (Ly6-G/C)-PerCP-Cy5.5 (1:300) (all from BD Biosciences) and NK1.1-brilliant violet 421 (1:300) E-cadherin-PE (1:300) (from BioLegend, San Diego, CA, USA). Cells were sorted (at low pressure with a 100-μm nozzle) on a MoFlo sorter (Beckman-Coulter, Brea, CA, USA). Cells were not cultured in the period between isolation and sorting.
Kurklu B., Whitehead R., Ong E., Minamoto T., Fox J., Mann J., Judd L., Giraud A, & Menheniott T. (2014). Lineage-specific RUNX3 hypomethylation marks the preneoplastic immune component of gastric cancer. Oncogene, 34(22), 2856-2866.
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4

Enrichment and Characterization of Splenic APCs

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Spleens were perfused with collagenase D (1 mg/ml) and DNAse I (0.1 mg/ml), cut into small pieces and incubated at 37°C for 45 min. Cell preparations were filtered using 100‐μM cell strainers, and APC populations were enriched using magnetic depletion of B, NK, and T cells (Miltenyi). For experiments that focused only on cDC, cDC were enriched using CD11c‐positive selection (Miltenyi). Cell suspensions were labeled with the following antibodies: CD3‐APC (145‐2C11, 1/300 eBioscience), CD19‐APC (ID3, 1/300 Biolegend), NK1.1‐APC (PK136, 1/300 BD Pharmingen), B220‐BV510 (RA3‐6B2, 1/500, BD Horizon), CD11b PE‐CF594 (M1/70, 1/3000 BD Horizon), CD11c PE‐Cy7 (HL3, 1/400 BD Pharmingen), Ly6C‐PerCP‐Cy5.5 (AL‐21, 1/1000 BD Pharmingen), CD64 BV421 (X54‐5/7.1, 1/300 Biolegend), CD8α APC‐Cy7 (53‐6.7, 1/200 BD Pharmingen), or XCR1‐PE (REA707, 1/100 Miltenyi). Cells were sorted on a BD AriaSorp (BD Biosciences) with purity routinely higher than 95%. Expression of MHC II was assessed in a separate mix since the anti‐I‐Ab AF700 (M5/114, 1/500 BD Pharmingen) is a blocking antibody. For experiments with Karma mice in which cDC1 are Tomato+, CD11b‐PerCP‐Cy5 (M1/70, 1/200 BD Pharmingen) was used. All flow cytometry samples were run on a Fortessa (BD Biosciences) and analyzed using FlowJo software.
Draheim M., Wlodarczyk M.F., Crozat K., Saliou J., Alayi T.D., Tomavo S., Hassan A., Salvioni A., Demarta‐Gatsi C., Sidney J., Sette A., Dalod M., Berry A., Silvie O, & Blanchard N. (2017). Profiling MHC II immunopeptidome of blood‐stage malaria reveals that cDC1 control the functionality of parasite‐specific CD4 T cells. EMBO Molecular Medicine, 9(11), 1605-1621.
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