Three pairs of tumor samples from Mettl1cKO‐Ctrl and Mettl1cKO mice were lysed into single‐cell suspension by the above method. The cells from mouse tumor were stained with the antibodies against CD45 (1:200, 11‐0451‐81, Thermo Fisher), CD11b (1:200, 48‐0112‐82, Thermo Fisher), CD11c (1:200, 25‐0114‐81, Thermo Fisher), F4/80 (1:200, 45‐4801‐80, Thermo Fisher), and CD326 (1:200, 17‐5791‐80, Thermo Fisher) for 20 min at 25°C, followed by cleaning with staining buffer. Intracellular staining (CD206, 1:200, 12‐2061‐80, Thermo Fisher) was performed using Intracellular FIX/PERM kit (Thermo Fisher). Data acquisition was performed on Attune NxT4 (Thermo Fisher) and analyzed via FlowJo.
Intracellular fix perm kit
Manufactured by Thermo Fisher Scientific
The Intracellular FIX/PERM kit is a laboratory product designed for the fixation and permeabilization of cells. It facilitates the detection and analysis of intracellular proteins and other cellular components using flow cytometry or other analytical techniques.
Automatically generated - may contain errors
Lab products found in correlation
Cd11b, by Thermo Fisher Scientific (1 mentions)
F4 80, by Thermo Fisher Scientific (1 mentions)
Cd11c, by Thermo Fisher Scientific (1 mentions)
Cd206, by Thermo Fisher Scientific (1 mentions)
Cd326, by Thermo Fisher Scientific (1 mentions)
Celltrace violet dye, by Thermo Fisher Scientific (1 mentions)
Protein transport inhibitor cocktail, by Thermo Fisher Scientific (1 mentions)
Cd3 fitc, by Thermo Fisher Scientific (1 mentions)
Cd8 percp cy5, by Thermo Fisher Scientific (1 mentions)
Cd4 apc, by Thermo Fisher Scientific (1 mentions)
Macsquant16 instrument, by Miltenyi Biotec (1 mentions)
2 protocols using intracellular fix perm kit
1
Immunophenotyping of Mouse Tumor Cells
2
Quantifying Antigen-Specific T-Cell Responses
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Splenocytes from vaccinated and control mice were prepared and stained with CellTrace Violet dye according to the manufacturers instructions (ThermoFisher Scientific, MA, USA). One million splenocytes were then stimulated or left untreated for 4 days in wells of a flat bottom 96 well plate in duplicate. After 4 days, the cells were restimulated with antigen for a further 24 h. At 6–8 h prior to harvesting, protein transport inhibitor cocktail was added to the cells (ThermoFisher Scientific). The cells were harvested and the duplicate wells pooled into V bottom 96 well plates to reduce cell loss during the intracellular staining. The cell samples were then stained with T-cell surface markers (CD3 FITC, CD8 PerCp-Cy5.5, CD4 APC, all ThermoFisher Scientific) followed by intracellular staining of IFN-gamma (IFN-γ PE clone XMG1.2. ThermoFisher Scientific) using the intracellular fix/perm kit (ThermoFisher Scientific) according to the manufacturer’s instructions. The samples was analyzed with a MACSQuant16 instrument (Miltenyi Biotech).
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