Co-staining of KCa3.1 channels in NSCLC cells with a small-molecule fluorescent probe and antibody-based indirect immunofluorescence.

Co-staining of KCa3.1 channels in NSCLC cells with a small-molecule fluorescent probe and antibody-based indirect immunofluorescence. ChemMedChem. 2020 Oct 12;: Authors: Wünsch B, Brömmel K, Maskri S, Bulk E, Pethő Z, Rieke M, Budde T, Koch O, Schwab A Abstract The Ca 2+  activated potassium channel 3.1 (K Ca 3.1) is involved in critical steps of the metastatic cascade, such as proliferation, migration, invasion and extravasation. Therefore, a fast and efficient protocol for imaging of K Ca 3.1 channels was envisaged. The fluorescently labeled small molecule imaging probes 1 and 2 were synthesized by connecting a dimethylpyrrole-based BODIPY dye with a derivative of the K Ca 3.1 channel inhibitor senicapoc via linkers of different length. Patch-clamp experiments revealed the inhibition of K Ca 3.1 channels by the probes confirming interaction with the channel. Both probes 1 and 2 were able to stain K Ca 3.1 channels in non-small-cell lung cancer (NSCLC) cells following a simple, fast and efficient protocol. Pre-incubation with unlabeled senicapoc removed the punctate staining pattern showing the specificity of the probes 1 and 2 . Staining of the channel with the fluorescent probes 1 or 2 or with antibody-based indirect immunofluorescence yielded identical or very similar densities of stained K Ca 3.1 channels. However, co-staining using both methods did not lead to the expected overlapping punctate staining pattern. This observa...
Source: ChemMedChem - Category: Chemistry Authors: Tags: ChemMedChem Source Type: research