Samples were centrifuged at 8000× g for 30 min, and the supernatant was taken and loaded onto a Sephadex G75 (2.5 cm d × 50 cm l), previously equilibrated with Tris 150 mM at pH 8. Chromatography was run at a constant flux rate (2 mL/min and eluting fractions were characterized by SDS-PAGE to determine the fractions containing bands corresponding to the molecular weight of Cav-2α. These fractions were combined and dialyzed several times against 5 L of Tris 5 mM at pH 8 and concentrated using a Vivaspin Turbo 15 (3 kDa cut off) (Sartorious; Gloucestershire, UK).
Do you have any questions about this protocol?
Post your question to gather feedback from the community. We will also invite the authors of this article to respond.
Tips for asking effective questions
+ Description
Write a detailed description. Include all information that will help others answer your question including experimental processes, conditions, and relevant images.