Technical Reference #1743

1743.

Ca2+ Waves Initiate Antigen-Stimulated Ca2+ Responses in Mast Cells Roy Cohen; Alexis Torres; Hong-Tao Ma; David Holowka; Barbara Baird, Cornell University, Journal of Immunology, 183(1743), (2009)
Link To Paper

Materials & Methods:
The GCaMP2 construct (24) was provided by Dr. M. Kotlikoff Cornell University College of Veterinary Medicine. Small hairpin RNA (shRNA) plasmids targeting TRPCs (TRPC1 TRPC3 TRPC5 TRPC7 and GFP control) were characterized in RBL cells as previously described (25).Fluo-4 AM and Fluo-5F AM were purchased from Invitrogen/Molecular Probes. U73122 D-sphingosine apsigargin A23187 2-aminoethyldiphenyl borate (2-APB) and GdCl3 were from Sigma-Aldrich. NN -dimethylsphingosine (DM-sphingosine) was from Avanti Polar Lipids.

Microscopic Technique
Confocal Microscopy

Cell Type(s)
RBL-2H3 cells