PBMCs were mixed gently with anti-CD4-FITC and anti-CD25-APC, and anti-CD8-APC antibodies (BD Pharmingen™, USA) respectively and incubated for 30 min at 4 °C, and washed twice. Then, fixed, permeabilized, and stained with anti-Foxp3-PE, anti-IFNγ-PE-CY7, anti-IL4-APC-CY7, anti-IL17-BV421, anti-TNFα-Percpcy5.5, anti-Ki67-FITC, anti-Granzyme B-PE, and anti-TGFβ-APC (BD Pharmingen™, USA) for analysis of T cells. The samples were performed by flow cytometry on a FACS Aric III flow cytometer (BD Biosciences, USA). Flow cytometry data were analyzed using FlowJo software 10.4.
Anti granzyme b pe
Manufactured by BD
Sourced in United States
Anti-Granzyme B-PE is a fluorescently labeled antibody that binds to Granzyme B, a serine protease enzyme found in the cytotoxic granules of natural killer cells and cytotoxic T lymphocytes. The PE (phycoerythrin) fluorescent label allows for the detection and quantification of Granzyme B-positive cells using flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Anti ki67 fitc, by BD (2 mentions)
Anti cd4 fitc, by BD (2 mentions)
Anti foxp3 pe, by BD (1 mentions)
Anti tnfα percpcy5, by BD (1 mentions)
Anti tgfβ apc, by BD (1 mentions)
Anti ifn γ pe cy7, by BD (1 mentions)
Anti cd8 apc, by BD (1 mentions)
Anti cd25 apc, by BD (1 mentions)
Ionomycin, by Merck Group (1 mentions)
Phorbol myristate acetate (pma), by Merck Group (1 mentions)
Red blood cell lysis solution, by Qiagen (1 mentions)
Cd25 apc cy7, by BD (1 mentions)
Ifn γ pe cy7, by BD (1 mentions)
Fixation permeabilization solution kit, by BD (1 mentions)
Gata3 apc, by BD (1 mentions)
Il 4 apc cy7, by BD (1 mentions)
Tnfα apc cy7, by BD (1 mentions)
Foxp3 pe cy7, by BD (1 mentions)
Gr 1 v450, by BD (1 mentions)
Cd8a v450, by BD (1 mentions)
5 protocols using anti granzyme b pe
1
Multiparametric Flow Cytometry for T Cell Profiling
2
Modulating PBMC Cytolytic Function
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PBMCs were co-cultured with different concentrations of rIL-10 and/or rTGF-β for 5 h. Then, cells were stimulated with PMA (10 ng/μL) and ionomycin (1 μg/μL) (Sigma Chemical St. Louis, MO, USA) for 6 h at 37 °C. PBMCs were collected, and surface-stained with anti-CD3-PerCP and anti-CD56-PE-Cy7. Anti-granzyme B-PE and anti-perforin-fluorescein isothiocyanate (FITC) (BD Biosciences, San Jose, CA, USA) were used for intracellular staining, and cells detected by flow cytometry (LSR II).
3
Tumor Immune Profiling by Flow Cytometry
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Tumor xenografts were cut into pieces and digested with collagenase IV (Sigma, USA) and DNAse I (Sigma, USA) for 30 min at 37°C. Then, cell suspensions were passed through a 70-μm strainer to remove undigested tissues. Erythrocytes were removed by Red Blood Cell Lysis Solution (Qiagen, USA). For cell surface staining, 1 × 106 cells were incubated with anti-Fc receptor blocking antibody at 4°C for 30 min. For intracellular staining, 1 × 106 cells were fixed and permeabilized using the Fixation/Permeabilization Solution Kit (BD Bioscience, USA) according to the manufacturer’s instructions. Then, cells were stained with the indicated antibodies for 30 min at 4°C. Flow cytometry was performed using a BD Influx cell sorter (BD Bioscience). Flow cytometry data were analyzed by FlowJo version 10 (FlowJo, USA). The flow cytometry antibodies used in our study were CD45 APC, CD4 PE, CD8a V450, CD25 APC-CY7, FoxP3 PE-CY7, IFN-γ PE-CY7, IL-4 APC-CY7, Gr-1 V450, Ly6C FITC, CD11 b PE, F4/80 APC-CY7, Tim-3 PE-CY7, PD-1 PerCP CY5.5, T-bet FITC, GATA3 APC, TNF-α APC-CY7, anti-Ki67 FITC, and anti-Granzyme B PE, all from BD Bioscience.
4
Tumor-Infiltrating Immune Cell Profiling
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Tumor tissue was harvested and cut into small fragments followed by digestion with tumor disassociation kit for 30 min (Miltenyi Biotec, USA), and then filtered by 70 µm cell strainers. Mononuclear cells were enriched by percoll gradient centrifuging of the single cell suspension. Cells were washed with PBS and stained with Live/Dead dye, anti-CD4 FITC, anti-CD8 PE-Cy7, anti-Ki67 APC, anti-IFNγ PE-CF594, anti-FoxP3 Percp and anti-granzyme B PE antibodies, along with appropriate isotype controls (all from BD) for flow cytometry analysis (BD FACSCalibur).
5
Immune Checkpoint Modulation in Cancer
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Anti-human PD-L1 antibody avelumab was purchased from MedChemExpress and this experiment was carried out with a standard dose of 20 μg/ml. Pomalidomide was purchased from MedChemExpress and this experiment was carried out with the standard dose of 20 μg/ml. The anti-human PD-L1 antibody avelumab and the immunomodulatory drug Pomalidomide were dissolved in dimethylsulfoxide (DMSO) and stored at -20℃.
Anti-PD-L1-phycoerythrin (PE) (no. 561787), anti-CD73-PB450 (no. 561260), anti-CD90-fluorescein isothiocyanate (FITC) (no. 555462), anti-CD105-allophycocyanin (APC) (no. 562408), anti-CD34-peridinin chlorophyll protein (PerCP) (no. 555821) and anti-CD45-APC-A750 (no. 557833) were purchased from BD Pharmingen (Franklin Lakes, NJ, USA).
Anti-CD3-PerCP (no. 6610008), anti-CD4-FITC (no. 35-0048), anti-CD8-PE (no. 6610025), anti-CD56-PE (no. Z6410020) and anti-PD-1-APC (no.6610897) were obtained from Beijing Tongsheng shidai Biotech Co., Ltd. Anti-CD34-FITC (no. 340430), anti-perforin-PE, anti-granzyme B-PE and isotype controls were obtained from BD Biosciences (San Jose, CA, USA).
Anti-PD-L1-phycoerythrin (PE) (no. 561787), anti-CD73-PB450 (no. 561260), anti-CD90-fluorescein isothiocyanate (FITC) (no. 555462), anti-CD105-allophycocyanin (APC) (no. 562408), anti-CD34-peridinin chlorophyll protein (PerCP) (no. 555821) and anti-CD45-APC-A750 (no. 557833) were purchased from BD Pharmingen (Franklin Lakes, NJ, USA).
Anti-CD3-PerCP (no. 6610008), anti-CD4-FITC (no. 35-0048), anti-CD8-PE (no. 6610025), anti-CD56-PE (no. Z6410020) and anti-PD-1-APC (no.6610897) were obtained from Beijing Tongsheng shidai Biotech Co., Ltd. Anti-CD34-FITC (no. 340430), anti-perforin-PE, anti-granzyme B-PE and isotype controls were obtained from BD Biosciences (San Jose, CA, USA).
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