Immunomagnetic separation is a suitable method for electrophysiology and ion channel pharmacology studies on T cells.

Immunomagnetic separation is a suitable method for electrophysiology and ion channel pharmacology studies on T cells. Channels (Austin). 2021 Dec;15(1):53-66 Authors: Tajti G, Szanto TG, Csoti A, Racz G, Evaristo C, Hajdu P, Panyi G Abstract Ion channels play pivotal role in the physiological and pathological function of immune cells. As immune cells represent a functionally diverse population, subtype-specific functional studies, such as single-cell electrophysiology require proper subset identification and separation. Magnetic-activated cell sorting (MACS) techniques provide an alternative to fluorescence-activated cell sorting (FACS), however, the potential impact of MACS-related beads on the biophysical and pharmacological properties of the ion channels were not studied yet. We studied the aforementioned properties of the voltage-gated Kv1.3 K+ channel in activated CD4+ T-cells as well as the membrane capacitance using whole-cell patch-clamp following immunomagnetic positive separation, using the REAlease® kit. This kit allows three experimental configurations: bead-bound configuration, bead-free configuration following the removal of magnetic beads, and the label-free configuration following removal of CD4 recognizing antibody fragments. As controls, we used FACS separation as well as immunomagnetic negative selection. The membrane capacitance and of the biophysical parameters of Kv1.3 gating, voltage-dependence of steady-stat...
Source: Channels - Category: Molecular Biology Tags: Channels (Austin) Source Type: research