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Anti cd8 pe

Manufactured by Miltenyi Biotec
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Anti-CD8 PE is a laboratory reagent used for the detection and identification of CD8+ T cells in flow cytometry applications. It consists of a fluorescent phycoerythrin (PE) dye conjugated to an antibody that specifically binds to the CD8 surface marker on human cells.

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Lab products found in correlation

Anti cd4 fitc, by Miltenyi Biotec (3 mentions) Facscanto 2, by BD (3 mentions) Facsdiva software, by BD (2 mentions) Anti c45r b220 apc, by Miltenyi Biotec (2 mentions) Anti nk 1.1 percp vio770, by Miltenyi Biotec (2 mentions) Isotype matched control igg, by Miltenyi Biotec (2 mentions) Facscalibur, by BD (1 mentions) Anti cd3 fitc, by Miltenyi Biotec (1 mentions) Anti cd16 cd32 antibody, by BioLegend (1 mentions) Cellquest pro software, by BD (1 mentions) Facsverse, by BD (1 mentions) Facssuite, by BD (1 mentions) Anti cd4 pe, by Miltenyi Biotec (1 mentions) Anti cd19 fitc, by Miltenyi Biotec (1 mentions) Anti cd25 apc, by Miltenyi Biotec (1 mentions) Anti cd44 apc, by Miltenyi Biotec (1 mentions) Anti cd62l pe, by Miltenyi Biotec (1 mentions) Foxp3 staining buffer set, by Miltenyi Biotec (1 mentions) Anti cd3e, by BD (1 mentions) Qiazol lysis reagent, by Qiagen (1 mentions)

Spelling variants of this product

CD8-PE (11 citations) Anti-CD8 (8 citations) Anti-CD8-PE-Vio770 (3 citations)

5 protocols using anti cd8 pe

1

Flow Cytometry Analysis of Immune Cells

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Unpurified splenocytes or isolated subpopulations were subjected to flow cytometric analyses to validate the quality of the cells. Prior to staining, the Fc receptors were blocked by preincubation with anti‐CD16/CD32 antibodies (Biolegend, San Diego, CA, USA) for 10 minutes on ice. Surface staining was accomplished by incubating the cells with fluorochrome‐conjugated specific antibodies for 20 minutes in the dark on ice. The following antibodies (all purchased from Miltenyi Biotec) were employed: anti‐CD3‐FITC (130‐102‐496), anti‐CD4‐FITC (130‐102‐541), anti‐CD4‐PE (130‐102‐619), anti‐CD8‐PE (130‐102‐595), anti‐CD19‐FITC (130‐092‐042), anti‐CD25‐APC (130‐102‐787), anti‐CD44‐APC (130‐102‐563), anti‐CD62L‐PE (130‐102‐907).
Intracellular staining of FoxP3 was performed using an anti‐FoxP3‐PE antibody (130‐098‐119; Miltenyi Biotec) and the FoxP3 Staining Buffer Set (130‐093‐142; Miltenyi Biotec) following the given instructions.
Flow cytometric analyses were run on a FACS Verse (BD Biosciences) or FACS Calibur (BD Biosciences). A total of 10 000 events per sample were acquired and data were evaluated using the FACS Suite or CellQuest Pro software (both BD Biosciences).
Ehlers L., Rohde S., Ibrahim S, & Jaster R. (2018). Adoptive transfer of CD3+ T cells and CD4+ CD44high memory T cells induces autoimmune pancreatitis in MRL/MpJ mice. Journal of Cellular and Molecular Medicine, 22(4), 2404-2412.
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2

Phenotypic Analysis of T Cell Subsets

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The surface antigens CD4, CD8, CD3E, and TCR alpha were stained with the following fluorescent labeled antibodies: anti CD4-PE (Cat# 555347, BD Biscience), anti CD8-PE (Cat# 130-091-084, Miltenyi Biotech), anti-CD3E (BD Biosciences, Cat# 561806) and anti TCRA alpha/beta (Thermo Fisher Scientific, Cat# 17-9986-41) for 30 min at 4 °C. Cells were fixed in 1% paraformaldehyde and analyzed by flow cytometry (FACS Canto II, BD Biosciences).
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

Isolation and Activation of Immune Cells

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Blood was drawn into sodium heparin tubes (APP Pharmaceuticals, NDC 63323-540-11). CD4+, CD8+, and CD14+ cells were isolated from whole blood using anti-CD4/CD8/CD14 microbeads (130-090-877/130-090-878/130-090-879, Miltenyi Biotec) and Automatics purification as per the manufacturer’s instructions. A small portion of the cell isolate was set aside for flow cytometry using anti-CD3 APC-H7, anti-CD4 PE-Cy7, anti-CD11b-APC, anti-CD8 PE, anti CD14 – FITC (Miltenyi Biotec) to assess the purity. All antibodies were purchased from BD Pharmingen unless otherwise indicated. Isolated cells were re-suspended at 1 million/ml in RPMI containing 10% FBS and either immediately lysed in 0.7 ml QIAzol lysis reagent (Qiagen) or incubated for 4 h at 37 °C with 25 μl/ml anti-CD3/antiCD28 Human T-Activator Dynabeads (111.32D, Invitrogen) for CD4 and CD8 cells or 1 μg/ml pI:C (528906, Calbiochem), LPS (L439, Sigma) CPG ODN2006, Invivogen) and PGN (Sigma, 77140) for CD14 cells. Lysates were stored at −80 °C until processing for RNAseq.
Wang L., Felts S.J., Van Keulen V.P., Scheid A.D., Block M.S., Markovic S.N., Pease L.R, & Zhang Y. (2018). Integrative genome-wide analysis of long noncoding RNAs in diverse immune cell types of melanoma patients. Cancer research, 78(15), 4411-4423.
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4

Characterization of Immune Cells in NOD Mice

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Mononuclear spleen cells and spinal cord infiltrating leukocytes were characterized by flow cytometry. Briefly, MOG-immunized NOD mice were sacrificed at different disease score and perfused with cold saline prior to organ collection, then a single cell suspension for each sample was prepared (Cavone et al., 2011; Cavone et al., 2015) and stained with anti-CD4 FITC, anti-CD8 PE, anti-C45R (B220) APC and anti-NK 1.1 PerCP-Vio770 or isotype-matched control IgG antibodies (all from Miltenyi Biotec, Bergisch Gladbach, Germany) according to the manufacturer's instructions. The cells were then analyzed using a FACSCanto II flow cytometer (BD Biosciences) equipped with FACSDiva software (BD Biosciences), acquiring a total of 10 4 events for spleen extracts and at least 10 5 events for spinal cord extracts. 10 week-old NOD mice were used as control.
Urru M., Ranieri G., Mencucci R, & Chiarugi A. (2020). Comparison of the Anti-Inflammatory and Cytotoxic Potential of Different Corticosteroid Eye Drop Preparations. Ocular immunology and inflammation, 28(5).
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5

Characterization of Immune Cells in NOD Mice

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Mononuclear spleen cells and spinal cord infiltrating leukocytes were characterized by flow cytometry. Briefly, MOG-immunized NOD mice were sacrificed at different disease score and perfused with cold saline prior to organ collection, then a single cell suspension for each sample was prepared (Cavone et al., 2011; Cavone et al., 2015) and stained with anti-CD4 FITC, anti-CD8 PE, anti-C45R (B220) APC and anti-NK 1.1 PerCP-Vio770 or isotype-matched control IgG antibodies (all from Miltenyi Biotec, Bergisch Gladbach, Germany) according to the manufacturer's instructions. The cells were then analyzed using a FACSCanto II flow cytometer (BD Biosciences) equipped with FACSDiva software (BD Biosciences), acquiring a total of 10 4 events for spleen extracts and at least 10 5 events for spinal cord extracts. 10 week-old NOD mice were used as control.
Buonvicino D., Ranieri G., Pratesi S., Guasti D, & Chiarugi A. (2019). Neuroimmunological characterization of a mouse model of primary progressive experimental autoimmune encephalomyelitis and effects of immunosuppressive or neuroprotective strategies on disease evolution. Experimental neurology, 322.
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