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OMIP-048

Quantification of calcium sensors and channels expression in lymphocyte subsets by mass cytometry

Agnieszka Jaracz‐Ros, Patrice Hémon, Roman Krzysiek, Françoise Bachelerie, Géraldine Schlecht‐Louf, Hélène Gary‐Gouy Cytometry PART A, Volume 93, Issue 7, 681-684 (2018)

PURPOSE: The present mass cytometry panel 1, 2 was designed to simultaneously assess the ex vivo expression of 2 calcium (Ca2+)‐sensors and 4 Ca2+‐channels, the key components of the T cells “Ca2+‐toolkit,” in murine conventional (Tconv) and Foxp3+ regulatory (Treg) T‐cell subsets, including both naive and activated populations (Table 1), after a validation step of the selected antibodies (Abs, listed in Table 2) by flow cytometry.

CELL TYPE: Primary murine lymphocytes from spleen and lymph nodes

MACHINE: BD LSR Fortessa & Fluidigm CyTOF 1

Marker Target Clone Fluorochrome Purpose
DNA All Species Purified Nucleated cells
DNA All Species Purified Viability
CD45 Mouse 30-F11 Purified Lineage
CD3 epsilon Mouse 145-2C11 Purified Lineage
CD4 Mouse RM4-5 Purified Lineage
CD5 Mouse 53-7.3 Purified Lineage
CD8a Mouse 53-6.7 Purified Lineage
CD44 Mouse IM7 Purified Activation
CD62L Mouse Mel-14 Purified Activation
CD25 Mouse PC61 Purified Activation
CD45R Mouse RA3-6B2 Purified Lineage
CD19 Mouse 6D5 Purified Lineage
Ki-67 Mouse B56 Purified Proliferation
FoxP3 Mouse FJK-16s Purified Lineage
ORAI1 Mouse 3F6H5 Purified Ca2+-channel
ORAI2 Mouse Purified Ca2+-channel
ORAI3 Mouse Purified Ca2+-channel
STIM1 Mouse CDN3H4 Purified Ca2+-channel
STIM2 Mouse Purified Ca2+-sensor
VDAC1 Mouse 20B12AF2 Purified Ca2+-sensor/channel