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Using Alomone labs Tityustoxin-Kα-ATTO-594 in microscopy technique, Williams R.W. et al. showed recently that stimulating adenylate cyclase (AC) decreased surface KV1.2 within the pinceaus of cerebellar basket cell (BC) axon terminals3.
Tityustoxin-Kα-ATTO-594 labels KV1.2 channel in mouse cerebellum.A. Staining of free-floating mouse brain sections using Tityustoxin-Kα-ATTO-594 (#STT-360-AR) (red). Sections were then stained using Anti-KV1.2 (KCNA2) Antibody (#APC-010) (1:200) followed by goat anti-rabbit-AlexaFluor-488 (green). Tityustoxin-Kα-ATTO-594 and KV1.2 are both detected in the pinceau structures (yellow staining, arrow) of the cerebellum. B. Same brain sections as in A were labeled with Tityustoxin-Kα-ATTO-594 (red) followed by anti-Parvalbumin. Tityustoxin-Kα-ATTO-594 (red) labeled the pinceau structures (arrow) of the cerebellum. Parvalbumin stained not only the pinceau but also the soma of Purkinje cells. Co-localization between the two is depicted in yellow.Alomone Labs Tityustoxin-Kα-ATTO-594 binds KV1.2 including β2 transfected HEK293T cells and is displaced by unlabeled Tityustoxin-Kα.HEK293T cells transfected with KV1.2 and β2 were incubated with 200 nM Tityustoxin-Kα-ATTO-594 (#STT-360-AR) in the absence (A,B) and presence (C,D) of 5 µM Tityustoxin-Kα (#STT-360). Similar results were observed by using 100 nM Tityustoxin-Kα-ATTO-594 (data not shown).Alomone Labs Tityustoxin-Kα-ATTO-594 inhibits rKV1.2 heterologously expressed in Xenopus oocytes.A. Time course of KV1.2 channel currents at 60 mV before (black) and during (green) application of 100 nM Tityustoxin-Kα-ATTO-594 (#STT-360-AR) for 1 min (indicated as bar). Currents were elicited by application of voltage ramp from a holding potential of -80 mV to +60 mV in 100 ms. B. Example of superimposed current traces before (black) and during (green) 100 nM application of Tityustoxin-Kα-ATTO-594.
Tityustoxin Kα (#STT-360) is a potent and specific blocker of KV1.2 channels1,2. It was originally isolated from the scorpion Tityus serrulatus venom2.
The labeled version of the toxin, Tityustoxin-Kα-ATTO-594, has been tested in electrophysiology applications and is specially suited to experiments requiring simultaneous labeling of different markers.
Tityustoxin-Kα-ATTO-594 (#STT-360-AR) is a highly pure, synthetic, and biologically active conjugated peptide toxin.
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Alomone Labs Tityustoxin-Kα-ATTO-594 labels surface Kv1.2 channels in the cerebellar pinceaus of rat cerebellar slices.Overnight incubation of fixed rat cerebellar slices in 3 nM Tityustoxin Kα-ATTO-594 (#STT-360-AR) gives positive signal in the molecular layer and in the pinceaus of BC axon terminals. Scale bar, 10 µm.
Adapted from Figure 4 in Williams, M.R. et al.
- Williams, M.R. et al. (2012) J. Neurosci. 32, 9228.