Cd8 apc cy7

Manufactured by BioLegend

Sourced in United States, United Kingdom

CD8-APC-Cy7 is a fluorescent-conjugated monoclonal antibody that binds to the CD8 antigen, a cell surface glycoprotein expressed on a subset of T lymphocytes. It can be used in flow cytometry applications to identify and quantify CD8+ T cells within a sample.

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Lab products found in correlation

Close spelling variants of this product

Anti-CD8-APC-Cy7 (clone SK1; Anti-mouse CD8–APC-Cy7 APC/Cy7-conjugated anti-human CD8 mAb APC/Cy7-anti-CD8 mAb (clone 53–6.7, CD8-APC-Cy7 (clone SK1, Anti-CD8-APC-Cy7 APC-cy7 conjugated anti-mouse CD8 APC-Cy7 anti-human CD8 APC-Cy7-conjugated anti-CD8 CD8-APC

43 protocols using cd8 apc cy7

Immune Cell Profiling by Flow Cytometry

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Antibodies used in the studies are mentioned below:
Anti-mouse: CD3-Pacific Blue, CD4-PE, CD8-APCCy7, CD69-FITC, CD44-FITC, CD62L-APC, IFNγ-APC, IL-17-PECy7, CD11b-APCCy7, CD11c-APC, CD80-FITC, CD86-PerCPCy5.5, CD40-PE, CD4-APC, and CD4-FITC from Biolegend, USA.
Anti-mouse: p38, ph-p38 and β-Actin from Cell Signaling Technology.

Pahuja I., Verma A., Ghoshal A., Mukhopadhyay S., Kumari A., Shaji A., Chaturvedi S., Dwivedi V.P, & Bhaskar A. (2023). Biapenem, a Carbapenem Antibiotic, Elicits Mycobacteria Specific Immune Responses and Reduces the Recurrence of Tuberculosis. Microbiology Spectrum, 11(4), e00858-23.

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Tumor Infiltrating T Cell Analysis

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Mice receiving 5 × 10⁴ 4T1 or EMT6 cells were treated with antibodies as descried in the figures. Excised tumors were dissociated as a single cell using the gentleMACS Dissociator (Miltenui Biotec Inc., San Diego, CA, USA) with the mouse Tumor Dissociation kit (Miltenui Biotec) and lymphocytes were enriched on a Ficoll gradient (Sigma-Aldrich). T cells were isolated using Dynabeads untouched mouse T cell kit (Thermo Fisher Scientific). T cells were stained using anti-CD3-PerCP (BioLegend, San Diego, CA, USA), CD4-FITC (eBioscience, San Diego, CA, USA), CD8-APC/Cy7 (BioLegend), CD45.1-PE (BioLegend), and IFNγ-Pacific Blue antibodies. Stained samples were analyzed using a BD FACSCanto II (BD Bioscience) cytometer.

Li C.W., Lim S.O., Chung E.M., Kim Y.S., Park A.H., Yao J., Cha J.H., Xia W., Chan L.C., Kim T., Chang S.S., Lee H.H., Chou C.K., Liu Y.L., Yeh H.C., Perillo E.P., Dunn A.K., Kuo C.W., Khoo K.H., Hsu J.L., Wu Y., Hsu J.M., Yamaguchi H., Huang T.H., Sahin A.A., Hortobagyi G.N., Yoo S.S, & Hung M.C. (2018). Eradication of triple negative breast cancer cells by targeting glycosylated PD-L1. Cancer cell, 33(2), 187-201.e10.

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Atovaquone, Cabozantinib, and Nanoparticle Formulation

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Atovaquone (Ato), and cabozantinib (cabo) were purchased from Aladdin Reagent (Shanghai, China), while mPEG (2000)-TK-PLGA (2000) were bought from Ruixi Biotech (Shanxi, China). A cellular puncturing material borneol was purchased from Lanzhou Wotelaisi Biology (Gansu, China). ICG was purchased from Dandong Yichuang Pharmaceutical co., LTD (Dandong, China). FcR blocking reagents, CD11b-PE Vio770 was purchased from Miltenyi Biotec (Köln, Germany). Gr-1-Alexa Fluor 532 was bought from Novus Biological (Colorado, USA). CD11c-BB700 was purchased from BD Pharmingen (New Jersey, USA). CD3-Alexa Fluor 532 was bought from eBioscience (San Diego, USA). CD80-Percp/Cy5.5, F4/80-PE/Dazzle 549, MHC II-APC/Cy7, CD4-PE/Cy5 and CD8-APC/Cy7 were purchased from BioLegend (San Diego, USA). Dulbecco’s modified Eagle’s medium (DMEM), Roswell Park Memorial Institute 1640 (RPMI-1640) growth medium, fetal bovine serum (FBS) and penicillin–streptomycin solution were purchased from Gibco BRL (Maryland, USA).

Yang W., Pan X., Zhang P., Yang X., Guan H., Dou H, & Lu Q. (2023). Defeating Melanoma Through a Nano-Enabled Revision of Hypoxic and Immunosuppressive Tumor Microenvironment. International Journal of Nanomedicine, 18, 3711-3725.

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Multiparametric Flow Cytometry Immunophenotyping

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The BLCs and PBMCs were counted and assessed for viability using trypan blue (Sigma-Aldrich) and compound microscopy to ensure >90% lymphocyte viability. To assess distribution of CD4⁺ and CD8⁺ T cells, immune regulation and functionality of CD8⁺ T cells in HIV infection, cells from the two compartments were subjected to two panels of fluorescently labeled antibodies. Panel 1 (phenotypic panel): Live/dead-amcyan (Life Technologies, Carlsbad, CA, USA), CD3-BV650, CD4-BV711, CD14-APC-Cy7, PD-1-BV711, TIM-3-BV785 (BioLegend, San Diego, CA, USA), and CD8-PE Texas Red (Invitrogen, Carlsbad, CA, USA). Panel 2 (intracellular cytokine staining panel): Live/dead-amcyan (Life Technologies), CD3-Alexa700, CD4-BV711, CD8-APC-Cy7, granzyme b-Alexa647, IFNγ-Dazzle 549 (BioLegend). Panel 2 staining was done after stimulation with PMA/Ionomycin (25/500 ng/mL). Acquisition was performed on BD FACS Aria III. Flowjo v10.5 (Flowjo, LLC) was used for flow cytometry analysis.

Muema D.M., Mthembu M., Schiff A.E., Singh U., Corleis B., Chen D., Bassett T., Rasehlo S.S., Nyamande K., Khan D.F., Maharaj P., Mitha M., Suleman M., Mhlane Z., Naidoo T., Ramjit D., Karim F., Kwon D.S., Ndung'u T, & Wong E.B. (2020). Contrasting Inflammatory Signatures in Peripheral Blood and Bronchoalveolar Cells Reveal Compartment-Specific Effects of HIV Infection. Frontiers in Immunology, 11, 864.

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Multiparametric Flow Cytometry Panel

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CD8 APC‐Cy7 (#301016, BioLegend), CD3 PE‐Cy7 (#300420, BioLegend), CD3 PerCPCy5.5 (#300430, BioLegend), CD4 PE (#317410, BioLegend), eBioscience™ Fixable Viability Dye eFluor™ 506 (#65‐0866‐18, Thermo), streptavidin Brilliant Blue 515 (#564453, BD), streptavidin Brilliant Violet 421 (#405225, BioLegend), streptavidin Brilliant Violet 711 (#563262, BD), streptavidin Brilliant Violet 786 (#563858, BD), streptavidin APC (#405243, BioLegend), streptavidin PE‐Dazz594 (#405248, BioLegend).

Brunk F., Moritz A., Nelde A., Bilich T., Casadei N., Fraschka S.A., Heitmann J.S., Hörber S., Peter A., Rammensee H., Singh H., Walz J., Maurer D, & Wagner C. (2021). SARS‐CoV‐2‐reactive T‐cell receptors isolated from convalescent COVID‐19 patients confer potent T‐cell effector function. European Journal of Immunology, 51(11), 2651-2664.

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Selective T Cell Depletion for Tumor Studies

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T cell subsets were depleted by intraperitoneally administering 400 μg of depleting antibody twice before prophylactic administration of MC38 TEVs and CT26 tumor challenge. CD4 T cells were depleted with anti-CD4 mAb (Clone GK1.5, Bio X Cell). CD8 T cells were depleted with anti-CD8α (Clone 2.43, Bio X Cell). CD4 and CD8 T cell depletion were confirmed using flow cytometry on the BD FACS CantoII (BD Biosciences) from the mouse spleen, lymph node, and peripheral blood. Antibodies for flow cytometry were CD3-APC (1:200, BioLegend, cat. no. 100236), CD8-APC-Cy7 (1:100, BioLegend, cat. no. 100714), CD4-BV510 (1:100, BioLegend, cat. no. 100449), CD11b-FITC (1:100, BioLegend, cat. no. 101206), CD19-FITC (1:100, BioLegend, cat. no. 115506), Nk1.1-FITC (1:100, BioLegend, cat. no. 108706), and CD45-PE (1:200, BioLegend, cat. no.103106).

Gates T.J., Wangmo D., Zhao X, & Subramanian S. (2023). Allogeneic tumor cell-derived extracellular vesicles stimulate CD8 T cell response in colorectal cancer. Molecular Therapy Oncolytics, 31, 100727.

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Multicolor Flow Cytometry of Brain Myeloid Cells

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BSLs purified from perfused brain tissue of moribund C57BL/6 mice on day 6 post-infection were stained with TCRβ-FITC, CD3ε-PE, CD4-PerCP, CD4-PerCP/Cy5.5, CD8-APC/Cy7, CD11b-APC, CD14-PE/Cy7, and F4/80-Brilliant Violet 421™ antibodies purchased from Biolegend (San Diego, CA) and fixable viability dye eFluor^®506 (eBioscience, San Diego, CA). An Amnis^® ImageStream^®X MKII (Amnis Merck-Millipore, Seattle, WA) equipped with five lasers (355, 405, 488, 561, and 642nm) was used for acquisition. A minimum of 30,000 events were collected at 40X magnification using the INSPIRE^® software (Amnis Merck-Millipore, Seattle, WA). Analysis was performed using IDEAS® 6.1 software (Amnis Merck-Millipore, Seattle, WA). Focused events were first gated using the Gradient RMS feature. Singlets were then gated by aggregate discrimination using aspect ratio and area. Analysis was then performed on gated live CD11b^highCD14⁺F480⁺TCRβ⁺CD3ε−CD4−CD8− cells.

Oakley M.S., Chorazeczewski J.K., Aleshnick M., Anantharaman V., Majam V., Chawla B., Myers T.G., Su Q., Okoth W.A., Takeda K., Akue A., KuKuruga M., Aravind L, & Kumar S. (2018). TCRβ-expressing macrophages induced by a pathogenic murine malaria correlate with parasite burden and enhanced phagocytic activity. PLoS ONE, 13(7), e0201043.

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Splenocyte Stimulation and Flow Cytometry

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Splenocytes were processed as described in the previous section and stimulated with RBD peptides for 5 hours at 37°C with protein transport inhibitor (Invitrogen) and anti-mouse CD107a-FITC antibody (BioLegend). Cell stimulation cocktail and R10, with protein transport inhibitor, were used as positive and negative controls, respectively. After stimulation, cells were stained with Live/Dead violet (Invitrogen) for viability. Anti-mouse CD4-BV510, CD8-APC-Cy7, CD44-A700, and CD62L-BV711 antibodies were used for surface staining and CD3e-PE-Cy5, IFN-γ-APC, and TNF-α-BV605 (all from BioLegend) were used for intracellular staining. The samples were run on an 18-color LSRII flow cytometer (BD Biosciences) and analyzed by FlowJo software.

Konrath K.M., Liaw K., Wu Y., Zhu X., Walker S.N., Xu Z., Schultheis K., Chokkalingam N., Chawla H., Du J., Tursi N.J., Moore A., Adolf-Bryfogle J., Purwar M., Reuschel E.L., Frase D., Sullivan M., Fry B., Maricic I., Andrade V.M., Iffland C., Crispin M., Broderick K.E., Humeau L.M., Patel A., Smith T.R., Pallesen J., Weiner D.B, & Kulp D.W. (2022). Nucleic acid delivery of immune-focused SARS-CoV-2 nanoparticles drives rapid and potent immunogenicity capable of single-dose protection. Cell Reports, 38(5), 110318.

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Multiparameter Flow Cytometry of PBMCs

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PBMC were stained with the following antibodies: TCRC Pacific Blue, CD4 V500, V follow (Biolegend, UK), MR1 APC (26.5) (Biolegend, UK), CD3 PerCP, CD161 APC, CD8 APC-Cy7, CD19 APC-Cy7 and CD20 Pacific Blue. All antibodies were from BD biosciences unless specified. The samples were stained for 15 minutes at room temperature, processed and then analyzed on a FACS Canto II machine with FACS Diva software evaluation.

Gao Y., Rae W., Ramakrishnan K.A., Barcenas-Morales G., Döffinger R., Eren E., Faust S.N., Ottensmeier C.H, & Williams A.P. (2016). Mucosal-Associated Invariant T (MAIT) Cells Are Impaired in Th17 Associated Primary and Secondary Immunodeficiencies. PLoS ONE, 11(5), e0155059.

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Comprehensive Immune Cell Profiling

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We used the following antibodies: CD4-PErCP (100538), CD8-APC/Cy7 (100713), CD69-FITC (104506), CD25-APC (101910), IFNγ-APC (505810), IL17-PE/Cy7 (506922), CD11b-APC/Cy7 (101226), CD11c-APC (117310), CD80-FITC (104706), MHCII-PE (107607), CD3-PacificBlue (100214), CD4-PE (100512), CD4-APC (100516) and CD4-FITC (100510) from Biolegend, USA.
SIRT2 (ab211033), β-actin (ab8227), acH3K18 (ab40888), α-tubulin (ab184613), acetyl-α-tubulin (ab179484), acetyl-NFκB p65 (ab19870), Alexa Fluor 647 (ab150075) and IgG-APC isotype control (ab232814) from Abcam.
ERK1/2 (9102), Phospho-ERK1/2 (9101), p-38 (9212) and phospho-p38 (4511) from Cell Signaling Technology.

Bhaskar A., Kumar S., Khan M.Z., Singh A., Dwivedi V.P, & Nandicoori V.K. (2020). Host sirtuin 2 as an immunotherapeutic target against tuberculosis. eLife, 9, e55415.

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