Cd8 bv510

Manufactured by BD

Sourced in United States

The CD8-BV510 is a fluorochrome-conjugated antibody that specifically binds to the CD8 surface antigen. It is designed for use in flow cytometry applications to identify and analyze CD8-positive cells.

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Lab products found in correlation

Cd3 percp cy5, by BD (4 mentions) Ifn γ pe, by BD (3 mentions) Facscalibur, by BD (3 mentions) Cd4 pe cf594, by BD (3 mentions) Tim 3 bv650, by BD (3 mentions) Ifn γ pe, by BioLegend (2 mentions) Cd45 fitc, by Thermo Fisher Scientific (2 mentions) Cd4 v450, by BD (2 mentions) Trustain fcx anti mouse cd16 32, by BioLegend (2 mentions) Apc h2kd, by BioLegend (2 mentions) Fitc nk1, by Thermo Fisher Scientific (2 mentions) Cxcr3 apc, by BD (2 mentions) Pe cy7 tim3, by BioLegend (2 mentions) Pd 1 pe, by BioLegend (2 mentions) Cd3 bv711, by BD (2 mentions) V450 cxcr4, by BD (2 mentions) Software v9, by FlowJo (2 mentions) Cd4 apc cy7, by BD (2 mentions) Cd11b bv510, by BD (2 mentions) Tigit pe cy7, by Thermo Fisher Scientific (2 mentions)

25 protocols using cd8 bv510

Flow Cytometry Analysis of Immune Cells

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Single-cell suspensions were blocked with mouse FcR blocking reagent (Miltenyi Biotec) for 10 min at 4 °C prior to surface staining. Cell viability was assessed by Fixable viability dye eFluor 520 (eBioscience) to exclude dead cells. The following anti-mouse antibodies were used: FITC-CD11b, PE-F4/80, APC-F4/80, FITC-CD45, PerCP-eFluor 710-MHC Class II (I-A/I-E), PerCP-eFluor 710-CD3, FITC-CD3e, FITC-CD4, APC-CD4, APC-CD8a, PE-PD-L2, PE-B7-H2, PE-B7-H3, PE-B7-H4, PE-TIM-4, PE-VISTA from eBioscience; APC-CD206, PE-PD-L1, APC-CD86, PE/Cy7 Ki-67 from Biolegend; V450-CD4, BV510-CD8, PE-IFN-γ from BD. The following anti-human antibodies were used: PerCP-eFluor 710-CD3, APC-CD4, APC-CD8a from eBioscience; FITC-CD14, PE-PD-L1, APC-CD163, APC-CD86, FITC-HLA-DR, PE-Cy7-CD163, PE-CD25, PE-IFN-γ from Biolegend; FITC-CD8, PE-IL-10 from BD. For intracellular staining, cells were fixed and permeabilized with the Fixation/Permeabilization solution kit (BD). All flow cytometry data was acquired on FACSCalibur or LSRFortessa (BD, San Jose, USA) and analyzed by FlowJo V10 (TreeStar, Ashland, USA).

Wen Z.F., Liu H., Gao R., Zhou M., Ma J., Zhang Y., Zhao J., Chen Y., Zhang T., Huang F., Pan N., Zhang J., Fox B.A., Hu H.M, & Wang L.X. (2018). Tumor cell-released autophagosomes (TRAPs) promote immunosuppression through induction of M2-like macrophages with increased expression of PD-L1. Journal for Immunotherapy of Cancer, 6, 151.

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Cytokine Profiling of Murine Splenocytes

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A total of 2×10⁶ murine splenocytes were incubated in a 96 U bottom well (Sarstedt) along with brefeldin A (1:1,000) and monensin A (1:1,000) at 37 °C for 5 hours. Stimulants were added at 2 µg and 40 MOI of peptides or adenovirus, respectively. After stimulation, samples were stored at 4°C overnight. Samples were then processed by centrifuging and resuspending pellets in murine Fc-block (BioLegend) at 4°C for 15 min. Extracellular proteins were then stained using the following panel: BV711 CD3 (BD Biosciences, #563123), BV510 CD8 (BD Biosciences, #563068), PE-CF594 CD4 (BD Biosciences, #562285), APC CD62L (eBioscience, #17-0621-81), AF700 CD44 (BioLegend, #103026) and v450 CCR7 (BD Biosciences, #560805). After staining, samples were fixed and permeabilized using a Fixation/Permeabilization kit (BD Biosciences) and following the manufacturer’s instructions. Intracellular proteins were then stained with the following panel: BV650 TNF-α (BioLegend, #506333), PE-Cy7 IL-2 (BioLegend, #503832), PE IFN-γ (BioLegend, #505808) Perp-eFluor-710 (Invitrogen, #46-1541-82) and FITC IL-6 (BioLegend, #503806). Samples were then run on a Fortessa flow cytometer (BD Bioscience) and data was analyzed with FlowJo (FlowJo V.10.8.2)

Feola S., Hamdan F., Russo S., Chiaro J., Fusciello M., Feodoroff M., Antignani G., D'Alessio F., Mölsä R., Stigzelius V., Bottega P., Pesonen S., Leusen J., Grönholm M, & Cerullo V. (2024). Novel peptide-based oncolytic vaccine for enhancement of adaptive antitumor immune response via co-engagement of innate Fcγ and Fcα receptors. Journal for Immunotherapy of Cancer, 12(3), e008342.

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Multiparameter Flow Cytometry Analysis

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The antibodies were TruStain FcX anti-mouse CD16/32 (BioLegend), APC-H2Kd (BioLegend), BV711-CD3 (BD Horizon), PE-CF594-CD4 (BD Horizon), FITC-NK1.1 (Invitrogen), PE-PD1 (BioLegend), APC-CXCR3 (BD Pharmigen), PE-CY7-TIM3 (BioLegend), BV510-CD8 (BD Horizon), and V450-CXCR4 (BD Horizon).
The data were acquired using BD LSRFortessa flow cytometer and analyzed using FlowJo software v9 (Ashland, OR).

Feola S., Chiaro J., Martins B., Russo S., Fusciello M., Ylösmäki E., Bonini C., Ruggiero E., Hamdan F., Feodoroff M., Antignani G., Viitala T., Pesonen S., Grönholm M., Branca R.M., Lehtiö J, & Cerullo V. (2022). A novel immunopeptidomic-based pipeline for the generation of personalized oncolytic cancer vaccines. eLife, 11, e71156.

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Multiparameter Flow Cytometry Panel

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The antibodies were: TruStain FcX™ anti-mouse CD16/32 (BioLegend), APC-H2Kd
(BioLegend), BV711-CD3 (BD Horizon), PE-CF594-CD4 (BD Horizon), FITC-NK1.1
(Invitrogen), PE-PD1(BioLegend), APC-CXCR3 (BD Pharmigen), PE-CY7-TIM3
(BioLegend), BV510-CD8 (BD Horizon), V450-CXCR4 (BD Horizon).
The data were acquired using BDLSRFORTESSA flow cytometer and analysed using FlowJo software v9 (Ashland, Oregon, USA).

Feola S., Chiaro J., Martins B., Russo S., Fusciello M., Ylösmäki E., Hamdan F., Feodoroff M., Antignani G., Viitala T., Pesonen S.K., Grönholm M., Branca R.M., Lehtiö J., & Cerullo V. (2021). A novel immunopeptidomic-based pipeline for the generation of personalized oncolytic cancer vaccines.

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Comprehensive Immune Profiling by Flow Cytometry

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1 × 10⁶ splenocytes were taken for antibody staining and downstream analysis. Specific monoclonal antibodies were purchased from eBioscience (Thermofisher Scientific, Waltham, MA, USA) or BD Biosciences (San Jose, CA, USA): CD4 APC-Cy7 (RM4-5), CD8 BV510 (53-6.7), CD11b BV510 (M170), CD11c PECy7 (HL3), CD19 PerCP-Cy5.5 (ID3), CD69 BV421 (H1.2F3), CD279 PE (PD1; J43), Gr-1 Alexa700 (Ly6G/Ly6C; RB6-8C5), MHC-II FITC (I-A/I-E; 2G9) and CD16/32 (2.4G2). H-2D^b restricted LCMV tetramer staining and peptide restimulation were performed as previously described [20 (link)]. All flow cytometry data were collected on a Fortessa X20 or Fortessa1 (BD Biosciences) and analyzed using FlowJo software (version 10, Flowjo LLC, Ashland, OR, USA).

Preston S.P., Allison C.C., Schaefer J., Clow W., Bader S.M., Collard S., Forsyth W.O., Clark M.P., Garnham A.L., Li-Wai-Suen C.S., Peiris T., Teale J., Mackiewicz L., Davidson S., Doerflinger M, & Pellegrini M. (2023). A necroptosis-independent function of RIPK3 promotes immune dysfunction and prevents control of chronic LCMV infection. Cell Death & Disease, 14(2), 123.

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Multiparametric Flow Cytometry of PBMCs

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PBMCs were incubated with directly conjugated antibodies for 30 min at 4°C. The cells were washed before flow cytometry analysis. Antibodies used included anti-human CD3-BV786, CD4-APC-Fire750, CD8-BV510, CD45RA-AF700, CD70-PE, PD-1-BV711, 2B4-FITC, CD160-AF488, TIM-3-BV650, CD95-PE-CY7 (BD Biosciences, San Diego, CA, USA), CCR7-BV421, HLA-DR-AF700, CD38-BV421, CD28-BV711, CD27-BV650 (BioLegend, San Diego, CA, USA), TIGIT-PE-Cy7, LAG-3-APC (Ebioscience, San Diego, CA, USA) and the corresponding isotype controls. Data acquisition was performed on an LSR Fortessa flow cytometer (BD Biosciences), and data was analyzed with FlowJo software (Tree Star, Ashland, OR, USA).

Wang D., Du J., Song Y., Wang B., Song R., Hao Y., Zeng Y., Xiao J., Zheng H., Zeng H., Zhao H, & Kong Y. (2020). CD70 contributes to age-associated T cell defects and overwhelming inflammatory responses. Aging (Albany NY), 12(12), 12032-12050.

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Multiparameter Flow Cytometry of PBMCs

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PBMCs were isolated from whole blood cells using Ficoll media purchased from Haoyang Bio-Manufacture CO., LTD (Tianjin, China). Staining fixable viability stain 780 (BD#565388) was added into the cell suspension at 1:1000, followed by an incubation with human Fcblock (BD#564220). Surface markers were stained using CD3-PerCP-Cy5.5 (BD#560835), CD4-PE-Cy7 (BD#560649), CD8-BV510 (BD#563256), CD25-PE (BD#557138), CD127-AF647 (BD#558598), CD45RA-APC (BD#550855) and CCR7-FITC (BD#560548). For intracellular staining, cells were fixed and permeablized using BD CytoFix/CytoPerm Kit (BD#554715) and then the cytokines were stained using IL-4-APC (BD#560671), IFN-γ-FITC (BD#554700), TNF-α-BV421 (BD#562783), and IL-17-PE (BD#560487), before the final flow cytometry assay using BD CantoII. Raw data of flow cytometry was analyzed by FlowJo software.

Jia R., Wang X., Liu P., Liang X., Ge Y., Tian H., Chang H., Zhou H., Zeng M, & Xu J. (2020). Mild Cytokine Elevation, Moderate CD4+ T Cell Response and Abundant Antibody Production in Children with COVID-19. Virologica Sinica, 35(6), 734-743.

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Flow Cytometry Analysis of Tumor-Derived Cells

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All antibodies used for flow cytometry were purchased from BD Pharmingen and included CD3–PerCP-Cy5.5 (Cat. number 561108), CD4-FITC (Cat. number 553047), CD8-APC (Cat. number 553035), CD8-BV510 (Cat. number 563068), CD45–APC-Cy7 (Cat. number 557659), CD45-PE (Cat. number 553081), CD90.2-FITC (Cat. number 553003), CD326 (EpCAM)-APC (Cat. number 563478), FoxP3-PE (Cat. number 560408), and PD-L1–PE (Cat. number 558091). One million cells were stained with 1 μg of each fluorochrome-conjugated antibody and analyzed using a FACS Canto II (Becton Dickinson). Cell surface staining was performed by incubating tumor-derived cell populations with selected antibodies on ice in the dark for 30 minutes in PBS and 0.5% FBS (HyClone GE Healthcare Life Sciences). Intracellular staining of FoxP3 was performed using a mouse FoxP3 fixation and permeabilization kit (BD Pharmingen, cat. number 560409). Non-viable cells were excluded from further flow analysis using a Live/Dead Fixable Violet Dead Cell Stain Kit (ThermoFisher, cat. number L34955). Data were analyzed using FlowJo version 10.1r7 (FlowJo, LLC).

Zhao F., Evans K., Xiao C., DeVito N., Theivanthiran B., Holtzhausen A., Siska P.J., Blobe G.C, & Hanks B.A. (2018). Stromal Fibroblasts Mediate Anti–PD-1 Resistance via MMP-9 and Dictate TGFβ Inhibitor Sequencing in Melanoma. Cancer immunology research, 6(12), 1459-1471.

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Immunophenotyping of Whole Blood and PBMCs

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For immunofluorescence staining of whole blood and PBMC, antibodies included CD45-ECD (Beckman Coulter, Indianapolis IN), CD3-e780, CD4-e450, CD8-AF700, CD15-FITC (eBioscience, San Diego, CA), CD45-FITC, CD4-PerCP-Cy5-5, CD14-PECy7, HLA-DR-APC, HLA-DR-BV421, CD8-APC-H7, CCR7-BV605, CD25-BV605, CD45RA-PECy7, CD45RO-APC-H7, CCR4-PECy7, CD38-APC, CD127-BV650 (BD Bioscience, San Jose, CA), CD14-AF700, and CD3-AF700 (Biolegend, San Diego, CA). For phosphoSTAT staining, antibodies included CD3-BV785 (Biolegend), CD4-BV605, CD8-BV510, CD14-PECy7, CD19-BV421, CD16-BV650, pSTAT3-647, pSTAT5-PE (BD Biosciences), and pSTAT1-488 (Cell Signaling Technologies, Danvers, MA). Cell types were identified according to the criteria of the Human Immunology Project [19 (link)], and a full list of antibodies used to identify individual cell types including those not reported here can be found in Additional file 2: Table S1. Hematological toxicity was graded according to NCI CTCAE v4.0 guidelines and is reported in Additional files 3 and 4: Tables S2 and S3.

Crocenzi T., Cottam B., Newell P., Wolf R.F., Hansen P.D., Hammill C., Solhjem M.C., To Y.Y., Greathouse A., Tormoen G., Jutric Z., Young K., Bahjat K.S., Gough M.J, & Crittenden M.R. (2016). A hypofractionated radiation regimen avoids the lymphopenia associated with neoadjuvant chemoradiation therapy of borderline resectable and locally advanced pancreatic adenocarcinoma. Journal for Immunotherapy of Cancer, 4, 45.

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Quantification of Intracellular IL-17A in PBMCs

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PBMCs were stimulated by Cell Stimulation and Protein Transport Inhibitor Cocktail (2 μl/ml, Biogems, USA) for 4 h at 37 °C. Then, PBMCs were washed twice with phosphate buffer saline (PBS) and incubated with CD3-PerCP-Cy5.5 and CD8-BV510 (BD Biosciences, USA) for 20 min at room temperature. Cells were then washed twice with PBS, and fixed with Intracellular Fixation Buffer (eBioscience, USA) for 30 min at 4 °C. After fixation, cells were permeabilized with diluted Permeabilization Buffer (1 × , eBioscience, USA) for 20 min at room temperature. Cells were then washed twice and incubated with IL-17A-PE (eBioscience, USA) for 30 min at room temperature. Cells were then washed twice and resuspended and acquired on BD FACS Canto II flow cytometer.

Wang L., Luo Y., Li X., Li Y., Xia Y., He T., Huang Y., Xu Y., Yang Z., Ling J., Weng R., Zhu X., Qi Z, & Yang J. (2022). Talaromyces marneffei Infections in 8 Chinese Children with Inborn Errors of Immunity. Mycopathologia, 187(5-6), 455-467.

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