Transduced T cells were used in this experiment. The T cells were washed with phosphate-buffered saline (PBS), centrifuged, and then blocked with BSA. The cells were subsequently washed with PBS. Primary antibodies were added to the test samples according to the manufacturer’s recommendations. The cells in both groups were rinsed with PBS. After surface staining and to perform intracellular staining of BLIMP1, initially the cells were fixed and permeabilized with a True-NuclearTM Buffer Set (BioLegend) according to the manufacturer’s instructions and then stained with rat anti-human BLIMP1 antibody or rat IgG2a antibody (BD Biosciences, San Jose, CA, USA). Finally, the cells were assessed by flow cytometry (BD FACSCalibur or BD FACSCanto, BD Biosciences) and analyzed with FlowJo v. 10 software.
True nuclear buffer set
Manufactured by BioLegend
Sourced in United States
The True-NuclearTM Buffer Set is a collection of solutions designed for the isolation and analysis of nuclei from various cell types. The set includes buffers for cell lysis, nuclei enrichment, and nuclear staining, enabling researchers to extract and study the nuclear components of their samples.
Automatically generated - may contain errors
Lab products found in correlation
Facscalibur, by BD (1 mentions)
Facscanto, by BD (1 mentions)
Anti cd4 percp clone sk3, by BD (1 mentions)
Esp3i enzyme, by Thermo Fisher Scientific (1 mentions)
Dynabead human t activator cd3 cd28, by Thermo Fisher Scientific (1 mentions)
Anti cd8 pe clone hit8a, by BD (1 mentions)
Anti cd8 pacific blue clone sk1, by BioLegend (1 mentions)
Primescript rt reagent kit, by Takara Bio (1 mentions)
Sybr green master mix, by Thermo Fisher Scientific (1 mentions)
Turbofect transfection reagent, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Rpmi 1640 medium, by Merck Group (1 mentions)
L glutamine, by Merck Group (1 mentions)
Rneasy micro kit, by Qiagen (1 mentions)
2 protocols using true nuclear buffer set
1
Intracellular Staining of BLIMP1 in T Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
Transduced T cells were used in this experiment. The T cells were washed with phosphate-buffered saline (PBS), centrifuged, and then blocked with BSA. The cells were subsequently washed with PBS. Primary antibodies were added to the test samples according to the manufacturer’s recommendations. The cells in both groups were rinsed with PBS. After surface staining and to perform intracellular staining of BLIMP1, initially the cells were fixed and permeabilized with a True-NuclearTM Buffer Set (BioLegend) according to the manufacturer’s instructions and then stained with rat anti-human BLIMP1 antibody or rat IgG2a antibody (BD Biosciences, San Jose, CA, USA). Finally, the cells were assessed by flow cytometry (BD FACSCalibur or BD FACSCanto, BD Biosciences) and analyzed with FlowJo v. 10 software.
+ Expand
Transduced T cells were used in this experiment. The T cells were washed with phosphate-buffered saline (PBS), centrifuged, and then blocked with BSA. The cells were subsequently washed with PBS. Primary antibodies were added to the test samples according to the manufacturer’s recommendations. The cells in both groups were rinsed with PBS. After surface staining and to perform intracellular staining of BLIMP1, initially the cells were fixed and permeabilized with a True-NuclearTM Buffer Set (BioLegend) according to the manufacturer’s instructions and then stained with rat anti-human BLIMP1 antibody or rat IgG2a antibody (BD Biosciences, San Jose, CA, USA). Finally, the cells were assessed by flow cytometry (BD FACSCalibur or BD FACSCanto, BD Biosciences) and analyzed with FlowJo v. 10 software.
2
T-Cell Activation and Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
Esp3I enzyme (#FD0454), TurboFect Transfection Reagent, and Dynabead Human T-Activator CD3/CD28 (#11131D) were purchased from Thermo Fisher Scientific. RPMI-1640 medium, fetal bovine serum, and L-glutamine were purchased from Sigma-Aldrich. Lymphodex (#002041500) was obtained from Inno-train, the RNeasy Micro Kit (#74004) from Qiagen, PrimeScript RT Reagent Kit from Takara, SYBR Green Master Mix (#4367659) from Applied Biosystems Life Technologies, Inc., True-NuclearTM Buffer Set from BioLegend, Rat IgG2a antibody from BD Biosciences, and human IL-2 from Royan Institute. The anti-CD3-FITC (clone HIT3a, #555339), anti-CD4-PerCP (clone SK3, #566316), anti-CD8-PE (clone HIT8a, #555635), anti-CD45RA-PE (clone 5H9, #556627), anti-CD45RA-APC (clone 5H9, #561210), and anti-BLIMP1-PE (clone 6D3, #564702) antibodies from BD Biosciences, the anti-CD8-Pacific Blue (clone SK1, #344718) and anti-CD62L-FITC (clone DREG-56, #304812) antibodies from BioLegend, the anti-CD3-APC-Alexa Flouor 750 (clone UCHT1, #A66329), anti-CD4-Krome Orange (clone 13B8.2, #A96417) antibodies from Beckman Coulter, and the anti-CD107a-PE (clone eBioH4A3, #12-1079-42) antibody from eBioscience were used for the flow cytometry experiments.
+ Expand
Esp3I enzyme (#FD0454), TurboFect Transfection Reagent, and Dynabead Human T-Activator CD3/CD28 (#11131D) were purchased from Thermo Fisher Scientific. RPMI-1640 medium, fetal bovine serum, and L-glutamine were purchased from Sigma-Aldrich. Lymphodex (#002041500) was obtained from Inno-train, the RNeasy Micro Kit (#74004) from Qiagen, PrimeScript RT Reagent Kit from Takara, SYBR Green Master Mix (#4367659) from Applied Biosystems Life Technologies, Inc., True-NuclearTM Buffer Set from BioLegend, Rat IgG2a antibody from BD Biosciences, and human IL-2 from Royan Institute. The anti-CD3-FITC (clone HIT3a, #555339), anti-CD4-PerCP (clone SK3, #566316), anti-CD8-PE (clone HIT8a, #555635), anti-CD45RA-PE (clone 5H9, #556627), anti-CD45RA-APC (clone 5H9, #561210), and anti-BLIMP1-PE (clone 6D3, #564702) antibodies from BD Biosciences, the anti-CD8-Pacific Blue (clone SK1, #344718) and anti-CD62L-FITC (clone DREG-56, #304812) antibodies from BioLegend, the anti-CD3-APC-Alexa Flouor 750 (clone UCHT1, #A66329), anti-CD4-Krome Orange (clone 13B8.2, #A96417) antibodies from Beckman Coulter, and the anti-CD107a-PE (clone eBioH4A3, #12-1079-42) antibody from eBioscience were used for the flow cytometry experiments.
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