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2 protocols using anti plc β2

1

Endothelial Cell Culture Reagents

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All endothelial cell culture media were obtained from Lonza, Inc. (Walkersville, MD, USA). Dulbecco's modified Eagle's medium, fetal bovine serum, trypsin-EDTA and penicillin/streptomycin (antibiotics) were purchased from Hyclone Laboratories, Inc. (Logan, UT, USA). Anti-PLC-β1, anti-PLC-β2, anti-PLC-β3 and anti-PLC-β4 were obtained from Santa Cruz (California City, CA, USA). Anti-actin antibody was purchased from MP Biomedicals (Aurora, OH, USA). NG2 antibody was obtained from Millipore Bioscience (Temecula, CA, USA). Anti-pH3 antibody was purchased from Cell Signaling Technology (Boston, MA, USA). GSL I-isolectin B4 was obtained from Vector Laboratories (Burlingame, CA, USA). Alexa Fluor 405- and 488-conjugated streptavidin, and Alexa Fluor 488-conjugated goat anti-rabbit secondary antibodies were purchased from Molecular Probes, Inc. (Carlsbad, CA, USA). IRDye700- and IRDye800-conjugated rabbit and mouse secondary antibodies were obtained from Li-COR Bioscience (Lincoln, NE, USA).
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2

Antibody Characterization of Ion Channels

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The following primary antibodies were used: anti-PLCγ1 (Santa Cruz, sc-426) for immunocytochemistry and proximity ligation assay experiments; anti-PLCγ2 (Santa Cruz, sc-31751) for immunocytochemistry and proximity ligation assay experiments; anti-PLCβ2 (Santa Cruz, sc-206) for immunocytochemistry and proximity ligation assay experiments; anti-TRPC6 (Alomone, ACC-120) for immunocytochemistry, electrophysiology, and Ca2+ imaging experiments; anti-TRPC6 (Santa Cruz, sc-19197) for proximity ligation assay experiments; anti-TRPM4 (Abcam, ab63080) for immunocytochemistry and proximity ligation assay experiments; anti-TRPM4 (Santa Cruz, sc-67125) for immunocytochemistry and proximity ligation assay experiments; and anti-BKCa (Alomone, APC-021) for proximity ligation assay experiments. Secondary antibodies used were the following: sheep anti-rabbit Texas Red (Abcam, ab6793-1), bovine anti-rabbit FITC (Santa Cruz, sc-2365), donkey anti-goat Texas Red (Santa Cruz, sc-2783), and donkey anti-goat FITC (Abcam, ab6881).
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