Fresh parotid gland cell suspensions were incubated with antibodies (identified below) for 30 min at 4 °C and washed twice in PBS/0.5% BSA/2 mM EDTA. The following antibodies were used: anti-human-CD19-eF450 (clone HIB19), anti-human-CD27-APC (clone O323), both from eBioscience, and anti-human-FcRL4-PE (clone 413D12, Biolegend). Immediately before sorting, cells were stained with propidium iodide (eBioscience) for live/dead discrimination. Gating was performed as described in Supplementary Fig. 1 . Cells were sorted as 5 cells/well into 96-wells PCR plates containing 2 μl of lysis buffer (0.2% Triton X-100 (Sigma-Aldrich) + 2 U/μL RNAse inhibitor (Westburg-Clontech)), 1 μl of 10 μM oligo-dT30VN primer (Biolegio) and 1 μl of 4 × 10 mM dNTP mix (Westburg-Fermentas) per well. Cells were sorted on a MoFlo Astrios cell sorter (Beckman Coulter).
Anti human cd27 apc clone o323
Manufactured by Thermo Fisher Scientific
Anti-human-CD27-APC (clone O323) is a monoclonal antibody conjugated with the fluorescent dye Allophycocyanin (APC). It is designed to recognize the CD27 antigen, which is expressed on the surface of various human immune cells, including T cells, B cells, and natural killer cells. This product can be used for the identification and characterization of CD27-positive cells in flow cytometry applications.
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2 protocols using anti human cd27 apc clone o323
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Parotid Gland Cell Immunophenotyping
2
Parotid Gland Cell Immunophenotyping
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Fresh parotid gland cell suspensions were incubated with antibodies (identified below) for 30 min at 4 °C and washed twice in PBS/0.5% BSA/2 mM EDTA. The following antibodies were used: anti-human-CD19-eF450 (clone HIB19), anti-human-CD27-APC (clone O323), both from eBioscience, and anti-human-FcRL4-PE (clone 413D12, Biolegend). Immediately before sorting, cells were stained with propidium iodide (eBioscience) for live/dead discrimination. Gating was performed as described in Supplementary Fig. 1 . Cells were sorted as 5 cells/well into 96-wells PCR plates containing 2 μl of lysis buffer (0.2% Triton X-100 (Sigma-Aldrich) + 2 U/μL RNAse inhibitor (Westburg-Clontech)), 1 μl of 10 μM oligo-dT30VN primer (Biolegio) and 1 μl of 4 × 10 mM dNTP mix (Westburg-Fermentas) per well. Cells were sorted on a MoFlo Astrios cell sorter (Beckman Coulter).
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