T cell population detection post vaccination was assessed using flow cytometry assay and 50 μL whole EDTA blood. Red blood cells were lysed with ACK buffer and washed twice with 1% (v/v) FBS in 1× PBS. The fluorophore-conjugated antibody mixture was added to the sample and incubated on ice for 20 min. FITC anti-mouse CD3ε Antibody (Cat#10036), APC/Cyanine7 anti-mouse CD8a Antibody (Cat#100714), APC anti-mouse CD4 Antibody (Cat#100412), and PE anti-mouse CD19 Antibody (Cat#115508) were bought from Biolegend (San Diego, CA, USA). Cells were then resuspended with 1% FBS in 1× PBS after washing 3 times. All samples were run on the flow cytometer.
Apc cyanine7 anti mouse cd8a antibody
Manufactured by BioLegend
Sourced in United States
The APC/Cyanine7 anti-mouse CD8a Antibody is a fluorescently-labeled antibody that binds to the CD8a surface marker on mouse cells. It is designed for use in flow cytometry applications to identify and analyze CD8a-positive cell populations.
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2 protocols using apc cyanine7 anti mouse cd8a antibody
1
T Cell Phenotyping Post Vaccination
2
Evaluating BMDC Stimulation of T Cells
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To study the secretion of the pro-inflammatory factor of BMDCs in vitro, LPS was used to stimulate the maturation of BMDCs after treatment with different BMDC formulations. The mRNA levels of TNF-α, IL-6, IL-2 were evaluated by q-PCR assays at 36 h after Ova uptake by BMDCs according to the standard protocol of q-PCR mentioned above.
For the evaluation of the antigen presentation ability, the specific tetramer-positive T cells after treatment with different DC formulations was detected as follows. Different DC formulations were incubated with naive OT-1 T cells isolated from the spleens of OT-1 mice for 48 h at a ratio of OT-1 T cells to DCs of 5 : 1. Then the cells were collected and stained by APC/ Cyanine7 anti-mouse CD8a antibody (100714, biolegend) and PE anti-mouse H-2Kb bound to SIINFEKL antibody (141603, biolegend) for flow cytometry assay.
The activation, proliferation, cytokine secretion, and cytotoxicity of OT-1 T cells after treatment with different DC formulations were measured to evaluate the stimulation of T cells by DCs. For the activation status of OT-1 T cells, the cells were
For the evaluation of the antigen presentation ability, the specific tetramer-positive T cells after treatment with different DC formulations was detected as follows. Different DC formulations were incubated with naive OT-1 T cells isolated from the spleens of OT-1 mice for 48 h at a ratio of OT-1 T cells to DCs of 5 : 1. Then the cells were collected and stained by APC/ Cyanine7 anti-mouse CD8a antibody (100714, biolegend) and PE anti-mouse H-2Kb bound to SIINFEKL antibody (141603, biolegend) for flow cytometry assay.
The activation, proliferation, cytokine secretion, and cytotoxicity of OT-1 T cells after treatment with different DC formulations were measured to evaluate the stimulation of T cells by DCs. For the activation status of OT-1 T cells, the cells were
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