T cell activation was assessed by flow cytometry using the following fluorescent-conjugated antibodies: CD69-Brilliant Violet 421 (BioLegend 310929, Dedham, MA), CD25-PerCP Cy5.5 (aka IL2RA BioLegend 302626), and HLA-DR-APC (BioLegend 307610). Cell cycle was assessed using the Click-It EdU Kit (Life Technologies, Carlsbad, California). A pulse of EdU was performed 1 hour at 37°C 4%CO2 prior to cell harvest and preserved for immunofluorescent staining with Cytofix fixation buffer for 15 minutes at 37°C (BD Biosciences, San Jose, California). Apoptosis was assessed using 7-AAD (2.5 ul) plus Alexa Fluor 647-conjugated Annexin-V (2.5 ul) in the presence of 1X Annexin-V staining buffer (Life Technologies). All flow cytometry experiments were completed using either the LSR II or Fortessa Flow Cytometer (BD biosciences).
Annexin 5 staining buffer
Manufactured by Thermo Fisher Scientific
The 1X Annexin-V Staining Buffer is a laboratory reagent designed for the detection and analysis of apoptosis. It provides the necessary environment for the binding of Annexin-V, a protein that specifically binds to phosphatidylserine, to cells undergoing apoptosis. This buffer ensures the appropriate conditions for the Annexin-V assay, which is a widely used method for evaluating programmed cell death.
Automatically generated - may contain errors
Lab products found in correlation
Brilliant violet 421 cd69, by BioLegend (2 mentions)
Hla dr apc, by BioLegend (2 mentions)
Cytofix fixation buffer, by BD (2 mentions)
Alexa fluor 647 conjugated annexin 5, by Thermo Fisher Scientific (2 mentions)
Click it edu kit, by Thermo Fisher Scientific (2 mentions)
Fortessa flow cytometer, by BD (2 mentions)
Cd25 percp cy5, by BioLegend (2 mentions)
2 protocols using annexin 5 staining buffer
1
Comprehensive T Cell Immunophenotyping
2
Comprehensive T Cell Immunophenotyping
Check if the same lab product or an alternative is used in the 5 most similar protocols
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T cell activation was assessed by flow cytometry using the following fluorescent-conjugated antibodies: CD69-Brilliant Violet 421 (BioLegend 310929, Dedham, MA), CD25-PerCP Cy5.5 (aka IL2RA BioLegend 302626), and HLA-DR-APC (BioLegend 307610). Cell cycle was assessed using the Click-It EdU Kit (Life Technologies, Carlsbad, California). A pulse of EdU was performed 1 hour at 37°C 4%CO2 prior to cell harvest and preserved for immunofluorescent staining with Cytofix fixation buffer for 15 minutes at 37°C (BD Biosciences, San Jose, California). Apoptosis was assessed using 7-AAD (2.5 ul) plus Alexa Fluor 647-conjugated Annexin-V (2.5 ul) in the presence of 1X Annexin-V staining buffer (Life Technologies). All flow cytometry experiments were completed using either the LSR II or Fortessa Flow Cytometer (BD biosciences).
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