Cd8 bv786

Manufactured by BD

Sourced in Germany

The CD8-BV786 is a fluorochrome-conjugated antibody used in flow cytometry applications to identify and analyze CD8+ T cells. It provides a specific and sensitive detection of the CD8 surface marker expressed on cytotoxic T lymphocytes.

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Lab products found in correlation

Cd4 buv395, by BD (3 mentions) Cd127 pe cy7, by BioLegend (2 mentions) Cd3 bv711, by BioLegend (2 mentions) Lsrfortessa, by BD (2 mentions) Cd3 fitc, by Beckman Coulter (2 mentions) Cd4 percp cyanine5, by BioLegend (2 mentions) Cd16 cd56 pe, by Beckman Coulter (2 mentions) Cd45 efluor506, by Thermo Fisher Scientific (2 mentions) Cd19 percp5, by BioLegend (2 mentions) Cd25 apc, by BioLegend (2 mentions) Ccr6 bv421, by BioLegend (2 mentions) Cd45ro ecd, by Beckman Coulter (2 mentions) Lsrfortessa cell analyzer, by BD (1 mentions) Il 2 pe, by BD (1 mentions) Cd44 apc cy7, by BD (1 mentions) Ifn γ pe dazzle594, by BioLegend (1 mentions) Live dead fixable aqua dead cell stain, by Thermo Fisher Scientific (1 mentions) Prism 6, by GraphPad (1 mentions) Cd27 apc efluor780, by Thermo Fisher Scientific (1 mentions) Cd28 pe cy7, by BD (1 mentions)

Spelling variants of this product

Anti-CD8-BV786 (11 citations) Anti-CD8 BV786 (RPA-T8; (2 citations)

13 protocols using cd8 bv786

Multicolor Flow Cytometry Staining

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Splenocytes were stained according to the manufacturer’s guidelines with CD4-BUV395, CD8-BV786, CD44-APC-Cy7, KLRG-1-V450, IL-2-PE (BD Pharmingen), LIVE/DEAD Fixable Aqua Dead Cell Stain (ThermoFisher Scientific), and CD-127-PE-Cy7, CD3-BV711, Thy1.1-A700, IFN-γ-PE/Dazzle594 (BioLegend). Flow cytometry was performed using a BD LSR Fortessa Cell Analyzer (BD Biosciences, San Jose, CA). Data collected were analyzed using FlowJo Software (Tree Star, San Carlos, CA) and analyzed with Prism 6 software (GraphPad Software Inc).

Bozeman A.M., Laurie S.J., Haridas D., Wagener M.E, & Ford M.L. (2018). Transplantation Preferentially Induces a KLRG-1lo CD127hi Differentiation Program in Antigen-Specific CD8+ T Cells. Transplant immunology, 50, 34-42.

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Multiparametric Immune Cell Profiling

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PBMCs were labeled with different combinations of the following antibodies: CD4 BUV737 (BD, clone SK3), CD8 BV786 (BD, clone RPA-T8), CD8 BUV805 (BD, clone SK1), CD3 V500 (BD, clone UCHT1), CD3 eVolve605 (eBioscience clone OKT3), CD27 APC-eFluor780 (eBioscience, clone), CD27 BV510 (Biolegend, clone O323), CD45RA BV650 (BD, clone HI100), CD45RA Qdot655 (Invitrogen, clone MEM-56), CD28 PE-Cy7 (BD, clone 28.2), CX3CR1 APC (eBioscience, clone 2A9-1), CCR7 BV421 (Biolegend, clone G043H7), CD127 BV421 (Biolegend, clone A019D5). Near-IR fixable dye (Invitrogen) was used to exclude dead cells from the analysis. For intracellular staining, the following antibodies were used: Hobit IgM (BD, clone Sanquin-Hobit/1), Eomes eFluor660 (eBioscience, clone WD1928), Tbet BV421 (Biolegend, clone 4B10), Granzyme B AF700 (BD, clone GB11), Perforin FITC (eBioscience, clone dG9), Perforin PE (eBioscience, clone B-D48), Granzyme K PE (Immunotools, clone 24C3), Granzyme K FITC (Immunotools, clone 24C3). To stain for Hobit IgM, a secondary anti-IgM labeled with PE or FITC was used. The cells were labeled according to manufacturer’s instructions. For the intracellular staining, the cells were fixed and permeabilized using the Foxp3/Transcription Factor Staining kit (eBioscience). The samples were measured in PBS 0.5% FCS with a LSR Fortessa (BD). The analysis was done using FlowJo Version 10 software.

Oja A.E., Vieira Braga F.A., Remmerswaal E.B., Kragten N.A., Hertoghs K.M., Zuo J., Moss P.A., van Lier R.A., van Gisbergen K.P, & Hombrink P. (2017). The Transcription Factor Hobit Identifies Human Cytotoxic CD4+ T Cells. Frontiers in Immunology, 8, 325.

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Comprehensive Immune Profiling by Flow Cytometry

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Whole blood was used to calculate absolute leukocyte counts by Trucount analysis (BD Biosciences) according to manufacturer’s protocol. The following antibodies were used: CD45-PerCP (BioLegend), and CD3-APC-R700, CD4-BV711, CD8-BV786, all purchased from BD Biosciences. Using the bead count of the Trucount tube and the CD3⁺ cell count, absolute cell numbers of the proliferation/apoptosis and senescence panel were also calculated. In addition, for all individuals, HLA typing on whole blood was performed by flow cytometry at the first visit. For all these HLA-types, we characterized the known immunodominant CMV epitopes for which commercial dextramers are available. Antibodies used were HLA-A1/36-biotin/strep-PE-CF594, HLA-A2-V450, HLA-A3-APC, HLA-A24-PE, HLA-B7-FITC, and HLA-B8-PE-Cy7. Cells were measured on a Fortessa flow cytometer (BD Biosciences) and analyzed with FlowJo V10 software.

van den Berg S.P., Derksen L.Y., Drylewicz J., Nanlohy N.M., Beckers L., Lanfermeijer J., Gessel S.N., Vos M., Otto S.A., de Boer R.J., Tesselaar K., Borghans J.A, & van Baarle D. (2021). Quantification of T-cell dynamics during latent cytomegalovirus infection in humans. PLoS Pathogens, 17(12), e1010152.

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Comprehensive Immune Profiling by Flow Cytometry

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All FCM studies were performed using single-cell suspensions, and cells were stained in accordance with the standard protocols. Antibodies recognizing CD45-eFluor506 (#69045942, Thermo), CD4-Percp/Cyanine5.5 (#30050, Biolegend), CD8-Bv786 (#563823, BD Pharmingen,), CD3-FITC, CD16/CD56-PE (#A07735, Beckman) and CD19-PerCP5 (#302227, Biolegend) were used. Samples were analysed using a flow cytometer (BD Biosciences), and subsequent analysis was performed using FlowJo 10.1 software.

Liao X., Chen L., Liu J., Hu H., Hou D., You R., Wang X, & Huang H. (2022). m6A RNA methylation regulators predict prognosis and indicate characteristics of tumour microenvironment infiltration in acute myeloid leukaemia. Epigenetics, 18(1), 2160134.

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Multiparametric Phenotyping of PBMCs

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Standard extracellular staining was performed on PBMCs using the following fluorophore-labeled antibodies: CD14/CD19-V500 (BD Pharmingen), (Biolegend), CD8-BV786 (BD Pharmingen), CD28-PerCP-Cy5.5 (Biolegend), CCR7-PE-Cy7 (Biolegend), CD25-APC (Biolegend), CD45RA-qDot655 (Life Technologies), and CD4-APC-H7 (BD Pharmingen).

Cortes-Cerisuelo M., Laurie S.J., Mathews D.V., Winterberg P.D., Larsen C.P., Adams A.B, & Ford M.L. (2017). Increased Pre-Transplant Frequency of CD28+ CD4+ TEM Predicts Belatacept-Resistant Rejection in Human Renal Transplant Recipients. American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons, 17(9), 2350-2362.

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FACS Analysis of CD8+ Subsets in Breast Cancer

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For the FACS analysis of the CD8 subsets, human breast cancer patients (n = 14) from the University Cancer Center (UCT) Frankfurt study (ICD10‐Code: C50.9) were analysed and as described elsewhere.¹⁷ The following FACS antibodies were used for CD8 gating: anti‐CD4‐PE‐CF594 (clone: RPA‐T4, RRID: AB_11154394), CD8‐BV786 (clone: RPA‐T8, RRID: AB_2687487), CD33‐BV510 (clone: WM53, RRID: AB_2738102), CD19‐APC‐H7 (clone: SJ25C1, AB_1645468) and TCRαβ‐FITC (clone: T10B9.1A‐31, RRID: AB_10892811) all from BD Biosciences (San Diego, CA) and anti‐CD45‐AF700 (clone: 2D1, RRID:AB_2566373), CD56‐PerCP‐Cy5.5 (clone: 5.1H11, RRID: AB_2563914) and anti‐CD279‐APC (PD‐1) (clone: EH12.2H7, RRID: AB_940473), all from BioLegend, San Diego, CA. The study was approved by the Institutional Review Boards of the UCT Frankfurt and the Ethics Committee at University Hospital Frankfurt (approval numbers: SGO‐01‐2014 and SGO‐01‐2016).

Burkard T., Herrero San Juan M., Dreis C., Kiprina A., Namgaladze D., Siebenbrodt K., Luger S., Foerch C., Pfeilschifter J.M., Weigert A, & Radeke H.H. (2022). Differential expression of CD8 defines phenotypically distinct cytotoxic T cells in cancer and multiple sclerosis. Clinical and Translational Medicine, 12(12), e1068.

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Quantifying CD4+ T-cell Proliferation Suppression

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Either ex vivo- or 7-day-cultured CD4⁺ T-cells were placed in flow cytometric suppression assays, as described previously [11 , 13 (link), 14 (link)]. Briefly, responder CD4⁺ T-cells were stained with CFSE, followed by culture 1 μg/ml of fixed anti-CD3 (eBiosciences, 16-0037-85) and 1μg/ml of fixed anti-CD28 (eBiosciences, 16-0289-85) in the presence or absence of ex vivo sorted autologous bulk CD8⁺ T-cells at 1:0, 1:1 and 1:0.5 CD4:CD8 cell concentrations. On day 7 of culture, cells were stained for anti-CD4 PE-Cy7 (BD, 557852), anti-CD3 AlexaFluor700 (BD, 557943), anti-CD8 Pacific Blue (Biolegend, 344718) and flow cytometrically assessed for CD4 proliferating fraction (CFSE dilution). Percent proliferation and percent suppression were calculated as described previously [13 (link)]. Other antibodies utilized include: CD45RO Pacific Blue, Biolegend 304216; CD45RO PE, Biolegend 304244; CD45RA Fitc, BD 555488; CD8 PE, BD 340046; CD8 BV786, BD 563823; CD4 BV786, BD 563877; CD4 APC, BD 561841; CD4 PE, BD 555347; CD3 APC, Biolegend 300412; CD25 PE, BD 555432; CD25 APC, BD 555434; CD25 PacBlue, Biolegend 356130. Cells were analyzed on a BD LSRII or Cytek Aurora. Data were analyzed with FlowJo software v10.

Crawford M.P., Borcherding N, & Karandikar N.J. (2023). IL-17 cytokines preferentially act on naïve CD4+ T cells with the IL-17AF heterodimer inducing the greatest functional changes. PLOS ONE, 18(4), e0285166.

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Isolation and Stimulation of OT-II Splenocytes

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Splenocytes were isolated from the spleens of OT-II mice, disaggregated using a mesh screen, and then treated to osmotically lyse red blood cells with an ammonium chloride/potassium chloride lysis buffer (0.15 M NH₄Cl, 10 mM KHCO₃, 0.1 mM EDTA). Splenocytes were then cultured in RPMI 1640 medium supplemented with L-glutamine, 10% fetal calf serum, 200 U/mL penicillin/streptomycin, 1% sodium pyruvate, 1% HEPES, and 50 µM β-MeOH with 2 µg/mL class-II OVA peptide (ISQAVHAAHAEINEAGR). TLR agonists were added 1 hour before stimulating with the peptide at the following concentrations: 10 µg/mL Poly(I:C) HMW, 5 µM ODN 1826. After 72 hours, cells were analyzed via flow cytometry with the following antibodies: CD3-FITC (BD 555274), CD4-BUV395 (BD 563790), CD8-BV786 (BD 563332), PD-1-PECF594 (BD 562523), 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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Identification of HIV-specific T follicular Helper Cells

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HIV-specific Tfh responses were defined using fluorochrome conjugated HLA class II tetramers. Briefly, cells were stained for 1 h at 37 °C with APC and PE conjugated HLA Class II tetramer complexes, washed in 2% FCS-PBS and then stained with these antibodies: LIVE/DEAD Fixable Blue dead cell stain kit (Thermo Fisher Scientific), CD3 BV711 (Biolegend), CD4 BV650 (BD Biosciences), CD8 BV786 (BD Biosciences), CXCR5 AF488 (BD Biosciences), CXCR3 BV605 (Biolegend), PD-1 BV421 (Biolegend) and CD45RA AF700 (Biolegend); for 20 min at RT. Cells were washed and acquired on the LSRFortessa (BD Biosciences).

Baiyegunhi O.O., Mann J., Khaba T., Nkosi T., Mbatha A., Ogunshola F., Chasara C., Ismail N., Ngubane T., Jajbhay I., Pansegrouw J., Dong K.L., Walker B.D., Ndung’u T, & Ndhlovu Z.M. (2022). CD8 lymphocytes mitigate HIV-1 persistence in lymph node follicular helper T cells during hyperacute-treated infection. Nature Communications, 13, 4041.

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Multiparameter Flow Cytometry Profiling of CAR T Cells

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Samples were spun down at 1300 RPM, then washed once with staining buffer. Samples collected from mice had 2 mL of Ack Lysis buffer added for 5 min to lyse red blood cells then washed once with PBS. Samples were stained for CAR surface expression for 30 minutes at room temperature with 100 μL solution containing the goat polyclonal anti-human IgG (Sigma-Aldrich, Cat# I1886–2ML) conjugated with Lightning-Link APC (Expedeon). After CAR staining, samples were washed three times with staining buffer and then incubated at 4°C for 30 minutes in 50 μL of an antibody cocktail to label human surface markers. The following anti-human antibodies were used: CD4 Pacific Blue (BioLegend Cat# 300521), CD45RA APCCy7 (BioLegend Cat# 304128), CD45RO BV570 (BioLegend Cat# 304226), CD62L BV605 (BD Biosciences Cat# 562719), CD95 PE (BioLegend Cat# 305608), CD8 BV786 (BD Biosciences Cat# 563823). Samples were washed three times and incubated with Live/Dead Aqua (Thermo-Fischer Scientific Cat# L-34966) for 10 minutes to discriminate live and dead cells. Samples were washed twice to remove Live/Dead Aqua before being run on a Fortessa.

Smole A., Benton A., Poussin M.A., Eiva M.A., Mezzanotte C., Camisa B., Greco B., Sharma P., Minutolo N.G., Gray F., Bear A.S., Baroja M.L., Cummins C., Xu C., Sanvito F., Goldgewicht A.L., Blanchard T., Rodriguez-Garcia A., Klichinsky M., Bonini C., June C.H., Posey AD J.r., Linette G.P., Carreno B.M., Casucci M, & Powell DJ J.r. (2022). Expression of Inducible Factors Reprograms CAR-T Cells for Enhanced Function and Safety. Cancer cell, 40(12), 1470-1487.e7.

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