naïve cd4 t cell isolation kit  (Miltenyi Biotec)

Journal: Immunity, Inflammation and Disease

Article Title: TRPM2 Deficiency Attenuates Allergic Rhinitis‐Like Inflammation With Altered Ca 2+ ‐NFAT Signaling, Treg Responses, and sIgE Production

doi: 10.1002/iid3.70490

Figure Lengend Snippet: TRPM2 deficiency is associated with attenuated Ca 2+ ‐NFAT signaling during T cell activation. (A) Intracellular Ca 2+ flux in splenic CD4 + T cells from WT and TRPM2 −/− mice, measured by flow cytometry using Fluo‐4 AM after stimulation with anti‐CD3/CD28 antibodies (5 μg/mL each). The graph shows the mean fluorescence intensity (MFI) over time from one representative experiment of three independent replicates. (B) Nuclear translocation of NFATc1 and IL‐2 production were assessed by Western blot from WT and TRPM2 −/− mice.

Article Snippet: Naïve CD4 + T cells were then isolated by negative selection using a commercial mouse CD4 + T Cell Isolation Kit (Cat. No. 130‐104‐454, Miltenyi Biotec, Bergisch Gladbach, Germany), according to the manufacturer's instructions.

Techniques: Activation Assay, Flow Cytometry, Fluorescence, Translocation Assay, Western Blot