Following in vitro re-stimulation, cells were surface-stained with fluorochrome-labeled antibodies anti-CD3-Alexa-Fluor700, anti-CD8-V500 and anti-CD4-APC Cy7 (BD Biosciences, BioLegend). In addition, HLA class I CMV tetramers-PE labeled (A*01 VTE, A*02 NLV, B*07 TPR, B*08 ELR and B*035 IPS) (Beckman Coulter or IBA Solution for Life Sciences), and Live/Dead Fixable Blue Dead-Cell Stain (Invitrogen) was used for gating on viable cells. In some experiments we used surface-stained with fluorochrome-labeled antibodies anti-PD1-FITC and anti CD160-PE Cy7 (BD Biosciences, BioLegend). Cytofix/Cytoperm (BD Biosciences) reagents were used to fix and permeabilize cells for intracellular cytokine staining (ICS) using anti-IFN-γ-PE Cy7 (or BV605), anti-IL-2-FITC, anti-Granzyme B-PE CF 594, anti-CD107a-Pacific Blue, anti-T-bet-PE or BV655 (BD Biosciences) and anti-Eomes-Alexa-Fluor 660 (eBiosciences).
Anti granzyme b pe cf 594
Manufactured by BD
Anti-Granzyme B-PE CF 594 is a fluorescently labeled antibody that binds to and detects Granzyme B, a protein involved in cytotoxic T-cell and natural killer cell-mediated apoptosis. The CF 594 fluorescent dye is conjugated to the antibody, allowing for detection and quantification of Granzyme B expression using flow cytometry or other fluorescence-based applications.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd4 apc cy7, by BD (2 mentions)
Cytofix cytoperm, by BD (2 mentions)
Anti cd8 v500, by BD (2 mentions)
Live dead fixable blue dead cell stain, by Thermo Fisher Scientific (2 mentions)
Anti cd107a pacific blue, by BD (2 mentions)
Anti cd3 alexafluor700, by BD (2 mentions)
Anti pd 1 fitc, by BD (1 mentions)
Anti il 2 fitc, by BD (1 mentions)
Anti ifn γ pe cy7, by BD (1 mentions)
Anti tnfα pecy7, by BD (1 mentions)
Anti t bet pe, by BD (1 mentions)
Live dead aqua, by Thermo Fisher Scientific (1 mentions)
Anti ccr7 pe cy7, by BD (1 mentions)
Anti cd8 percp, by BD (1 mentions)
Anti cd4 pacific blue, by BD (1 mentions)
Anti ki67 fitc, by BD (1 mentions)
Anti cd45ra apc cy7, by BioLegend (1 mentions)
Anti granzyme b apc, by BD (1 mentions)
Anti eomes efluor 660, by Thermo Fisher Scientific (1 mentions)
Anti tnf pacificblue, by BioLegend (1 mentions)
4 protocols using anti granzyme b pe cf 594
1
Comprehensive T-cell Phenotyping Protocol
2
Multiparametric Flow Cytometry for Immune Profiling
3
Multiparametric Flow Cytometry Analysis of T Cell Responses to TBEV
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T cell responses to TBEV were assessed using multi-color flow cytometry, and the monoclonal antibodies (mAbs) used were; anti-CD107a FITC, anti-CD4 Pacific Blue, anti-CD8 PerCP, anti-HLA-DR PerCP, anti-Ki67 FITC, anti-Ki67 Alexa Fluor 700, anti-Bcl2 PE, anti-CCR7 PE-Cy7, anti-MIP-1β PE, anti-CD14 BD horizon V500, anti-CD19 BD horizon V500, anti-perforin FITC and anti-granzyme B APC, anti-granzyme B PE-CF594 all from BD Biosciences (San Jose, CA). Anti-CD45RA APC-Cy7, anti TNF pacific blue, anti-IFN-γ Brilliant Violet 570, anti-CD27 Brilliant Violet 785, anti-CD27 Brilliant Violet 421, anti-Helios Pacific Blue, anti-T-bet Alexa Fluor 488, anti-T-bet PE-Cy7, anti-CCR7 Brilliant Violet 785, anti-CD279 Brilliant Violet 711, anti-CD27 biotin and anti-CD127 Brilliant Violet 570 were all from Biolegend (San Diego, CA). Anti-CD38 Alexa Fluor 700, anti-CD38 eFluor 650, anti-CD127 Alexa Fluor 780, anti-PD-1 (CD279) PE, anti-Eomes eFluor 660 and IgM eFluor 650 were all from eBioscience (San Diego, CA). Anti-CD4 Qdot 605, anti-CD8 Qdot705, anti-CD8 Qdot 605, Streptavidin-Qdot 585, anti-CD57 pure and Aqua Live/Dead were all from Invitrogen (Carlsbad, CA). Anti-CD3 ECD, anti-CD3 PE-Cy5, HLA-A2 CMV pp65 tetramer in PE and anti-CD56 ECD were from Beckman Coulter (Brea, CA). HLA-A2 ILLDNITTL tetramer in PE was kindly provided by the NIH Tetramer core facility.
4
Cytokine and Cytotoxic Molecule Profiling in Activated PBMCs
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Thawed PBMCs were incubated with Leukocyte Activation Cocktail with BD GolgiPlug (BD Bioscience) 2 μL/1 × 106 cells for 4 h. Cells were washed, fixed, and permeabilized with the BD IntraSure™ Kit and stained with anti-IFN-γ-FITC, anti-Perforin-BV421 and anti-Granzyme B-PE-CF594 (all from BD Biosciences) for 30 min before being detected by flow cytometry (BD FACSAria II, BD Bioscience). Data were analyzed with BD FACSDiva Software v7.0.
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