Name: Stichodactyla Toxin-ATTO Fluor-590
Brand: Alomone Labs
SKU: STS-400-AR

Overview

Cat #: STS-400-AR

Alternative Name K⁺ channel toxin ShK, Toxin κ-stichotoxin-She3a, Kappa-stichotoxin-She3a, κ-SHTX-She3a

Lyophilized Powder yes

Origin Modified synthetic peptide originated from Stichodactyla helianthus (Caribbean sea anemone).

MW: ~4627 Da

Purity: >99% (HPLC)

Form Lyophilized powder.

Effective concentration 1-100 nM.

Sequence RSCIDTIPKSRCTAFQCKHSMKYRLSFCRKTCGTC.

Modifications Cys³-Cys³⁵, Cys¹²-Cys²⁸, Cys¹⁷-Cys³².

Label ATTO-590. ATTO dyes are characterized by strong absorption (high extinction coefficient), high fluorescence quantum yield, and high photo-stability. ATTO-590 maximum absorption is 593 nm and maximum fluorescence is at 622 nm. The fluorescence is excited most efficiently in the 575-610 nm range. The extent of labeling is 1 molecule of dye per molecule ShK.

Structure

Activity Stichodactyla Toxin blocks K_V1.3, K_V1.1, K_V1.4, and K_V1.6 at subnanomolar concentrations and K_V3.2 channels at 1000-fold higher concentration than that required to inhibit K_V1.3 channels^1-3.

References-Activity

Pennington, M.W. et al. (1995) Int. J. Pept. Protein Res. 46, 354.
Beeton, C. et al. (2003) J. Biol. Chem. 278, 9928.
Middleton, R.E. et al. (2003) Biochemistry 42, 13698.

Accession number P29187.

Shipping and storage Shipped at room temperature. Product as supplied can be stored intact at room temperature for several weeks. For longer periods, it should be stored at -20°C.

Solubility Any aqueous buffer. Centrifuge all product preparations before use (10000 x g 5 min).

Storage of solutions Up to two weeks at 4°C or three months at -20°C.

Our bioassay

Live cell imaging of Stichodactyla Toxin-ATTO Fluor-590 in differentiated PC-12 cells.
Neurite outgrowth was induced in PC12 cells through 10 days exposure to 100 ng/ml Native mouse NGF 2.5S protein (>95%) (#N-100). (A,D) CellMask™ Actin 1X solution was applied for 30 minutes, resulting in a green fluorescence to visualize cellular membrane. (B,E) Following this, the same cells underwent incubation with 0.3 µM of Stichodactyla Toxin-ATTO Fluor-590 for 10 minutes at 37⁰C, followed by PBSX1 wash, leading to red fluorescence indicative of the distribution of K_V channels. (C,F) Live imaging of the differentiated PC-12 cells allowed observation of Stichodactyla toxin distribution among the cells.
Binding of Stichodactyla Toxin-ATTO Fluor-590 to Purkinje cells in the rat cerebellum.
Lightly-fixed floating rat brain sections were labeled with Stichodactyla Toxin-ATTO Fluor-590 (#STS-400-AR), (100 nM). A. Staining of Purkinje cells in the cerebellum (arrows). B. Staining of neurons in the deep cerebellar nuclei (arrows). DAPI is used as the counterstain (blue).
Direct flow cytometry of Stichodactyla Toxin in live intact rat PC-12 cells.
^___ PC-12 cells.
^___ PC-12 cells + 0.1 µM ShK (Stichodactyla Toxin) (#STS-400).
^___ PC-12 cells + 0.1 µM Stichodactyla Toxin-ATTO Fluor-590 (#STS-400-AR).
Direct flow cytometry of Stichodactyla Toxin in PC-12-derived extracellular vesicles (EVs).
^___ PC-12-derived EVs.
^___ PC-12-derived EVs + 50 nM ShK (Stichodactyla Toxin) (#STS-400).
^___ PC-12-derived EVs + 50 nM Stichodactyla Toxin-ATTO Fluor-590 (#STS-400-AR).
Alomone Labs Stichodactyla Toxin-ATTO Fluor-590 blocks K_V1.3 channels expressed in Xenopus oocytes.
A. Representative time course of Stichodactyla Toxin-ATTO Fluor-590 (#STS-400-AR) inhibition of K_V1.3 current. Membrane potential was held at -100 mV, current was elicited by a 100 ms voltage ramp to +60 mV every 10 sec, and significantly inhibited by 10 nM Stichodactyla Toxin-ATTO Fluor-590 (green). B. Superimposed traces of K_V1.3 current after application of control (black) and of 10 nM Stichodactyla Toxin-ATTO Fluor-590 (green), taken from the recording in A.

References - Scientific background

Castaneda, O. et al. (1995) Toxicon 33, 603.
Pennington, M.W. et al. (1995) Int. J. Pept. Protein Res. 46, 354.
Beeton, C. et al. (2003) J. Biol. Chem. 278, 9928.
Middleton, R.E. et al. (2003) Biochemistry 42, 13698.
Yan, L. et al. (2005) Mol. Pharmacol. 67, 1513.

Scientific background

ShK (Stichodactyla Toxin) is a peptide toxin originally isolated from the nematocyst of the sea anemone Stichodactyla helianthus¹.

ShK blocks K_V1.3, K_V1.1, K_V1.4, and K_V1.6 at subnanomolar concentrations and K_V3.2 channels at 1000-fold higher concentration than that required to inhibit K_V1.3 channels. It has been shown to block K_V current in DRG neurons, to displace radioactive Dendrotoxin from brain synaptosomes¹and inhibit I¹²⁵-Charybdotoxin binding to Jurkat T lymphocytes with an IC₅₀ of 32 pM^2,4.

Evidence also suggests that native K_V currents in the central nervous system, which are predominantly carried by K_V1.2 channels, are highly sensitive to this toxin⁵.

A fluorescently labeled synthetic ShK is used to recognize and to sort K_V1.3 containing lymphocytes^4,6.

Target K_V1.1, K_V1.3, K_V1.4, K_V1.6 channels

Peptide Content: 100%

Lyophilized Powder

Stichodactyla Toxin-ATTO Fluor-590 (#STS-400-AR) is a highly pure, synthetic, and biologically active peptide toxin conjugated to ATTO-590 fluorescent dye.

Benefits of Stichodactyla Toxin-ATTO Fluor-590:
✓ Localization and distribution
✓ Clustering and internalization kinetics
✓ Live cell imaging
✓ Single cell detection
✓ Binding kinetics
✓ Direct flow cytometry

We gladly take on collaboration projects. Please Contact Us.

Certificate of Analysis SDS

For research purposes only, not for human use

Last Update: 24/09/2024

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Stichodactyla Toxin-ATTO Fluor-590

Overview

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Scientific Background

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