CHO K1 cells were electroporated with pc3 sNHE-HA or with pc3 hHv1 using the Neon 100 Kit (Invitrogen, Carlsbad, USA) and a MicroPorator (Digital Bio) according to the manufacturer’s protocol (3 × 1650 mV pulses with a 10-ms pulse width). Cells were transferred into complete medium composed of F12 plus GlutaMAX (Invitrogen) and 10% fetal bovine serum (Biochrom, Berlin, Germany). To select monoclonal cells stably expressing SpSLC9C1 or hHv1, the antibiotic G418 (1200 mg ml−1; Invitrogen) was added to the cell culture medium 24 h after the electroporation. Monoclonal cell lines were identified by immunocytochemistry using a rat-anti-HA antibody (Roche Applied Science) or by electrophysiological recordings.
Neon 100 kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Neon 100 Kit is a laboratory equipment product by Thermo Fisher Scientific. It is designed for the study and analysis of various samples in a controlled laboratory environment. The kit includes essential components necessary for performing these tasks, but a detailed description of its core function is not available without the risk of bias or extrapolation.
Automatically generated - may contain errors
2 protocols using neon 100 kit
1
Stable Expression of SLC9C1 and Hv1 in CHO K1 Cells
2
Stable CHO K1 cells with cGMP sensor
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CHO K1 cells stably expressing a CNG-A2 channel-based cGMP sensor (8) were electroporated with pc6RhCG-mCherry using the Neon 100 kit (Invitrogen) and a MicroPorator (Digital Bio) according to the manufacturer's protocol (3 x 1,650 mV pulses with 10 ms pulse width). Cells were transferred into complete medium composed of F12 plus GlutaMax (Invitrogen) and 10% fetal bovine serum (Biochrom). For the selection of monoclonal cells stably expressing RhGC, the antibiotics G418 (400 µg/ml; Invitrogen) and Blasticidin (50 µg/µl; Invitrogen) were added 24 h after electroporation. Monoclonal cell lines were identified by fluorescence microscopy using mCherry expression.
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