Overview
Lyophilized Powder yes
Origin Synthetic peptide
MW: ~4900 Da
Purity: >98% (HPLC)
Effective concentration 0.5-50 nM.
Sequence VFINAKCRGSPECLPKCKECIGKAAGKCMNGKCKCYP.
Modifications Disulfide bonds between Cys7-Cys28, Cys13-Cys33 and Cys17-Cys35. Amino acid Alanine at position 20 was replaced by Cysteine (Bold in the sequence).
Label ATTO-594. Maximum absorption 601 nm; Maximum fluorescence 627 nm. The fluorescence is excited most efficiently in the 580 - 615 nm range. This label belongs to the class of Rhodamine dyes and can be used with fluorescent equipment typically optimized to detect Texas Red and Alexa-594.
Structure
Activity Tityustoxin-Kα blocks cloned KV1.2 with high potency1,2.
Using Alomone labs Tityustoxin-Kα-ATTO Fluor-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.
Using Alomone labs Tityustoxin-Kα-ATTO Fluor-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.
References-Activity
- Rogowski, R.S. et al. (1994) Proc. Natl. Acad. Sci. U.S.A. 91, 1475.
- Werkman, T.R. et al. (1993) Mol. Pharmacol. 44, 430.
- Williams, M.R. et al. (2012) J. Neurosci. 32, 9228.
Shipping and storage
The product is shipped as a lyophilized powder at room temperature. Upon receipt, store the product at -20°C. Protect from moisture. Avoid exposure to light.
Solubility The product is lyophilized in 0.5 ml conical vial.
Centrifuge the vial before adding solvent (10,000 x g for 5 minutes). The lyophilizate may be difficult to visualize. Add solvent directly to the centrifuged vial. Tap the vial to aid in dissolving the lyophilized product. Tilt and gently roll the liquid over the walls of the vial. Avoid vigorous vortexing. Light vortexing for up to 3 seconds is acceptable if needed.
Soluble in pure water at high-micromolar concentrations (50 µM - 1 mM). For long-term storage in solution, it is recommended to prepare a stock solution by dissolving the product in double distilled water (ddH2O) at a concentration between 100-1000x of the final working concentration. Divide the stock solution into small aliquots and store at -20°C. Before use, thaw the relevant vial(s) and dilute to the desired working concentration in your working buffer. Centrifuge all product preparations at 10,000 x g for 5 minutes before use. It is recommended to prepare fresh solutions in working buffers just before use. Avoid multiple freeze-thaw cycles to maintain biological activity. Avoid exposure to light.
Centrifuge the vial before adding solvent (10,000 x g for 5 minutes). The lyophilizate may be difficult to visualize. Add solvent directly to the centrifuged vial. Tap the vial to aid in dissolving the lyophilized product. Tilt and gently roll the liquid over the walls of the vial. Avoid vigorous vortexing. Light vortexing for up to 3 seconds is acceptable if needed.
Soluble in pure water at high-micromolar concentrations (50 µM - 1 mM). For long-term storage in solution, it is recommended to prepare a stock solution by dissolving the product in double distilled water (ddH2O) at a concentration between 100-1000x of the final working concentration. Divide the stock solution into small aliquots and store at -20°C. Before use, thaw the relevant vial(s) and dilute to the desired working concentration in your working buffer. Centrifuge all product preparations at 10,000 x g for 5 minutes before use. It is recommended to prepare fresh solutions in working buffers just before use. Avoid multiple freeze-thaw cycles to maintain biological activity. Avoid exposure to light.
Storage of solutions
Store the reconstituted solution at -20°C for the shortest time possible. Avoid multiple freeze-thaw cycles. We do not recommend storing the product in working solutions for longer than a day. Avoid exposure to light.
Our bioassay
Tityustoxin-Kα-ATTO Fluor-594 labels KV1.2 channel in mouse cerebellum.A. Staining of free-floating mouse brain sections using Tityustoxin-Kα-ATTO Fluor-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 Fluor-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 Fluor-594 (red) followed by anti-Parvalbumin. Tityustoxin-Kα-ATTO Fluor-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.
Live cell imaging of Tityustoxin-Kα-ATTO Fluor-594 in differentiated PC-12 cells.Neurite outgrowth was induced in PC12 cells through a weak exposure to 100 ng/ml Native mouse NGF 2.5S protein (>95%) (#N-100). (A) CellMask™ Actin 1X solution was applied for 30 minutes, resulting in a green fluorescence to visualize cellular membrane. (B) Following this, the same cells underwent incubation with 1 µM of Tityustoxin-Kα-ATTO Fluor-594 for 60 minutes at 370C, followed by PBSX1 wash, leading to red fluorescence indicative of the distribution of KV1.2 channels. (C) Live imaging of the differentiated PC-12 cells allowed observation of Tityustoxin-Kα distribution among the cells.
Direct flow cytometry of Tityustoxin-Kα in live intact rat PC-12 cells.___ PC-12 cells.
___ PC-12 cells + 1 µM Tityustoxin-Kα (#STT-360).
___ PC-12 cells + 1 µM Tityustoxin-Kα-ATTO Fluor-594 (#STT-360-AR).
Alomone Labs Tityustoxin-Kα-ATTO Fluor-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 Fluor-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 Fluor-594.
References - Scientific background
- Rogowski, R.S. et al. (1994) Proc. Natl. Acad. Sci. U.S.A. 91, 1475.
- Werkman, T.R. et al. (1993) Mol. Pharmacol. 44, 430.
- Williams, M.R. et al. (2012) J. Neurosci. 32, 9228.
Scientific background
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 Fluor-594, has been tested in electrophysiology applications and is specially suited to experiments requiring simultaneous labeling of different markers.
Target KV1.2 K+ channels
Peptide Content: 100%
Lyophilized Powder
Tityustoxin-Kα-ATTO Fluor-594 (#STT-360-AR) is a highly pure, synthetic, and biologically active conjugated peptide toxin.
✓ Localization and distribution
✓ Live cell imaging
✓ Single cell detection
✓ Direct flow cytometry
We gladly take on collaboration projects. Please Contact Us.
For research purposes only, not for human use
Last Update: 25/11/2024
Applications
Citations
Citations
Powered by BiozPublished figures using this product
Alomone Labs Tityustoxin-Kα-ATTO Fluor-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 Fluor-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.
Product citations
- Williams, M.R. et al. (2012) J. Neurosci. 32, 9228.
