- Protocol for dissection and culture of murine dorsal root ganglia neurons to study neuropeptide release.
Protocol for dissection and culture of murine dorsal root ganglia neurons to study neuropeptide release.
STAR protocols (2021-02-23)
Caroline Perner, Caroline L Sokol
PMID33615276
摘要
In this protocol, we provide step-by-step instructions for dissection and culture of primary murine dorsal root ganglia (DRG), which provide an opportunity to study the functional properties of peripheral sensory neurons in vitro. Further, we describe the analysis of neuropeptide release by ELISA as a possible downstream application. In addition, isolated DRGs can be used directly for immunofluorescence, flow cytometry, RNA sequencing or proteomic approaches, electrophysiology, and calcium imaging. For complete details on the use and execution of this protocol, please refer to Perner et al. (2020).
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Sigma-Aldrich
层粘连蛋白 来源于 Engelbreth-Holm-Swarm 小鼠肉瘤基底膜, 1-2 mg/mL in Tris-buffered saline, 0.2 μm filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
汉克斯平衡盐 溶液, Modified, with sodium bicarbonate, without phenol red, calcium chloride and magnesium sulfate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Glial Cell Line-derived Neurotrophic Factor from rat, recombinant, expressed in baculovirus infected Sf21 cells, lyophilized powder, suitable for cell culture, ≥97% (SDS-PAGE)