Stim1 antibody

Immunohistochemistry was used to detect the expression of Stim1 in all tissue specimens. The paraffin-embedded specimens were cut into 8-μm-thick sections, and deparaffinized and rehydrated in a graded series of alcohols. After antigen retrieval, the sections were incubated with 3% hydrogen peroxide for 10 min to block endogenous peroxidase activity, and then were immunostained with Stim1 antibody (1:50) (Santa Cruz Biotechnology, Inc., Paso Robles, CA, USA) at 4  °C overnight. After rinsing with PBS, the sections were incubated with horseradish peroxidase-conjugated secondary antibody (Zhongshan Jinqiao Co., Beijing, China) for 1 h. 3,3-diaminobenzidine tetrahydrochloride (DAB, Zhongshan Jinqiao Co.) was used as the substrate for the final visualization. Counterstaining was carried out with hematoxylin. The negative controls were processed in a same protocol with PBS instead of primary antibody. The sections were observed under a light microscope (Olympus, Tokyo, Japan).

Fura-2 AM was from Molecular Devices (San Jose, CA). Xestospongin C and rapamycin were from Cayman Chemical (Ann Arbor, MI). Bethanechol was from Alfa Aesar (Haverhill, MA). Apamin was from Tocris Bioscience (Bristol, UK). Stim1 antibody and mouse IgG-κ Fc binding protein conjugated to CFL 488 were from Santa Cruz Biotechnology (Dallas, TX). Secondary antibody (goat anti-mouse IgG conjugated to horseradish peroxidase) was from BioRad (Hercules, CA). Antibodies to phosphatidylinositol 4,5 bisphosphate (PIP₂) were from Echelon Biosciences (Salt Lake City, UT). All other reagents, unless otherwise indicated, were from Sigma-Aldrich (St. Louis, MO). Phalloidin conjugates were from Biotium (Fremont, CA). Plasmids encoding Pseudojanin (PJ) (Addgene plasmid # 37999), LYN11-FRB-CFP (Addgene plasmid # 38003), and PJ-DEAD (PJ-D) (Addgene plasmid # 38002) were gifts from Robin Irvine. GFP-C1-PLCdelta-PH was a gift of Tobias Meyer (Addgene plasmid # 21179).