Prior to labelling with conjugated antibodies, samples were incubated with Fc-block (BD Bioscience) for 10 min at RT. The following antibodies were used for phenotyping: PE-Cy5 anti-TCRαβ (LifeTechnologies), BV421 anti-TCRδ (BD Bioscience), APC-H7 anti-CD4 (BD Bioscience), PE anti-CD8α (BioLegend) and PE-Cy7 anti-CD8β (eBioscience). Before labelling with antibodies, cells were fixated and permeabelized using CytoFix/CytoPerm (BD Bioscience) according to instructions provided by the manufacturer. Forward and side scatters defined a lymphocyte gate without cell doublets. The T cell population comprises TCRαβ+ and TCRγδ+ cells, and each TCR type was defined as fractions of the T cell gate.
Anti cd8α pe
Manufactured by BioLegend
Sourced in United States
Anti-CD8α-PE is a fluorochrome-conjugated monoclonal antibody that binds to the CD8α antigen, which is expressed on the surface of cytotoxic T cells and a subset of natural killer cells. This product can be used for the identification and enumeration of CD8+ cells in flow cytometry applications.
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Lab products found in correlation
Anti cd4 apc h7, by BD (1 mentions)
Cytofix cytoperm, by BD (1 mentions)
Fc block, by BD (1 mentions)
Fitc anti cd11c clone n418, by BioLegend (1 mentions)
Anti cd80 pe, by BioLegend (1 mentions)
Anti cd16 32 mab clone 93, by BioLegend (1 mentions)
Anti cd86 pe, by BioLegend (1 mentions)
7mp two photon microscope, by Zeiss (1 mentions)
Hoechst 33342 dye, by Thermo Fisher Scientific (1 mentions)
Zen 2012, by Zeiss (1 mentions)
Annexin 5 fitc, by BioLegend (1 mentions)
Violet proliferation dye 450 vpd450, by BD (1 mentions)
Ghost dye red 780, by Cytek Biosciences (1 mentions)
Anti cd8α pe, by BD (1 mentions)
Anti cd8α alexafluor 647, by BD (1 mentions)
Anti ccr7 pe, by BD (1 mentions)
Anti cd8α apc, by BioLegend (1 mentions)
Anti cd45ra fitc, by BD (1 mentions)
Anti cd3 bv421, by BioLegend (1 mentions)
Anti cd8β apc, by Miltenyi Biotec (1 mentions)
5 protocols using anti cd8α pe
1
Phenotyping T Cell Subsets
2
Multicolor Flow Cytometry of Dendritic Cells
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DCs were preincubated with anti-CD16/32 mAb (clone 93, BioLegend), and were stained with the following mAbs: fluorescent isothiocyanate (FITC)-anti-CD11c (clone N418, BioLegend), FITC-anti-XCR1 (clone ZET, BioLegend), phycoerythrin (PE)-anti-CD11b (clone M1/70, BioLegend), PE-anti-CD8α (clone 53–6.7, BioLegend), PE-anti-Tim1 (clone RMT1-4, BioLegend), PE-anti-Tim3 (clone RMT3-23, BioLegend), PE-anti-Tim4 (clone RMT4-54, BioLegend), PE-anti-CD80 (clone 16-10A1, BioLegend), PE-anti-CD86 (clone GL-1, BioLegend), PE-Hamster IgG (clone eBio299Arm, eBioscience), PE-rat IgG2a (clone RTK2758, BioLegnd), PE-rat IgG2b (clone RTK4530, BioLegnd), PE/Cyanine7-anti-CD172a (clone P84, BioLegend), allophycocyanin (APC)-anti-CD8α (clone 53–6.7, BioLegend), APC-anti-Axl (clone MAXL8DS, Thermo Fischer Scientific), APC-anti-MerTK (clone 2B10C42, BioLegend).
3
In vivo Imaging of Intestinal CD8+ T Cells
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Mice were i.v. injected with 10 µg anti–CD8α-PE (BioLegend) 5 h before imaging. After anesthetization, mice were injected i.v. with Hoechst 33342 dye (Invitrogen), and the distal duodenum was exposed and opened along the antimesenteric border, as described previously (Edelblum et al., 2012 (link)). The mucosal surface was placed against a coverslip bottom of a 3D-printed cassette containing PBS. Images were acquired with ZEN2012 (Carl Zeiss) using a 7MP two-photon microscope (Carl Zeiss) equipped with a Chameleon laser (Coherent). Excitation wavelength was 880 nm. Images were acquired by taking 21-µm Z-stacks at 3-µm steps every 20 s. Each XY plane spans 512 × 512 µm2. Videos were made and analyzed with Imaris 7.4 ×64 (Bitplane).
4
Flow Cytometry Analysis of T Cell Subsets
5
Phenotyping of Cytotoxic T Cells
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Target cells were harvested and stained with fluorescent-labeled anti-human antibodies for 30 min at 4°C in the dark, as follows. The following antibodies were used: anti-CD8α-PE (BioLegend, San Diego, CA, USA; cat. no. 300908), anti-CD8α-APC-Cy7 (BioLegend; cat. no. 300926), anti-CD8β-APC (Miltenyi, Bergisch Gladbach, Germany; cat. no. 130-110-569), anti-CD3-BV421 (BioLegend; cat. no. 317344), CMV pp65495-503 Tetramer-PE (NLVPMVATV, ProImmune, Oxford, UK), and anti-4-1BB-APC (BioLegend; cat. no. 309810). Fluorescence was measured using a BD FACS Canto or Canto II (BD Biosciences, San Jose, CA, USA) and analyzed using FlowJo software (BD Biosciences).
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