Name: Tertiapin-Q-ATTO Fluor-488
Brand: Alomone Labs
SKU: STT-170-AG

Overview

Cat #: STT-170-AG

Lyophilized Powder yes

Origin Synthetic peptide

MW: 3195.5 Da

Purity: >95% (HPLC)

Effective concentration 50-200 nM.

Modifications Disulfide bonds between Cys³-Cys¹⁴, Cys⁵-Cys¹⁸. Lys²¹- C-terminal amidation.

Label ATTO-488. ATTO dyes are characterized by strong absorption (high extinction coefficient), high fluorescence quantum yield, and high photo-stability. ATTO-488 maximum absorption is 501 nm and maximum fluorescence is at 523 nm. The fluorescence is excited most efficiently in the 480 - 515 nm range. This label is analogous to the well known dye fluorescein isothiocyanate (FITC) and can be used with filters typically used to detect FITC. The extent of labeling is one molecule of dye per molecule of Tertiapin-Q.

Structure

Activity Tertiapin-Q blocks a range of inward rectifier K⁺ channels (K_ir), in particular ROMK1 and GIRK, but with no effect on K_ir2 family members^1,2. In addition, it was shown to inhibit acetylcholine induced K⁺ currents in the heart^3,4.

References-Activity

Jin, W. and Lu, Z. (1998) Biochemistry 37, 13291.
Jin, W. et al. (1999) Biochemistry 38, 14294.
Drici, M.D. et al. (2000) Br. J. Pharmacol. 131, 569.
Kitamura, H. et al. (2000) Pharmacol. Exp. Therap. 293, 196.

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. Avoid exposure to light.

Solubility Any aqueous buffer. Centrifuge all product preparations before use (10000 x g 5 min). Avoid exposure to light. The product is lyophilized in 0.5 ml conical vial.

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

Our bioassay

Double staining of GIRK2 channels in substantia nigra pars compacta using Tertiapin-Q-ATTO Fluor-488 and Guinea pig Anti-GIRK2 (K_ir3.2) Antibody.
Rat brain sections were first incubated with 33 nM Tertiapin-Q-ATTO Fluor-488 (#STT-170-AG), (green). The same sections were incubated with Guinea pig Anti-GIRK2 (K_ir3.2) Antibody (#APC-006-GP). A. Tertiapin-Q-ATTO Fluor-488 binding appears in profiles of substantia nigra pars compacta (SNC) neurons (arrows). B. GIRK2 staining (red) is detected in profiles of SNC neurons (arrows). C. Merge of the two images demonstrates co-staining of GIRK2 channels by Tertiapin-Q-ATTO Fluor-488 and by Guinea pig Anti-GIRK2 (K_ir3.2) Antibody. DAPI staining (blue) is used to stain nuclei.
Alomone Labs Tertiapin-Q-ATTO Fluor-488 binds GIRK1/4-Cherry transfected HEK293T cells.
Transfected cells were incubated in the presence of 200 nM Tertiapin-Q-ATTO Fluor-488 (#STT-170-AG). The labeled toxin accumulates on the membrane surface after 26 sec. No binding is achieved in untransfected cells (data not shown).
The pictures are a kind gift from the lab of Prof. Eithan Reuveny, Weizmann Institute, Israel.
Direct flow cytometry of Tertiapin-Q in live intact rat PC-12 cells.
^___ PC-12 cells.
^___ PC-12 cells + 1 µM Tertiapin-Q (#STT-170).
^___ PC-12 cells + 1 µM Tertiapin-Q-ATTO Fluor-488 (#STT-170-AG).
Alomone Labs Tertiapin-Q-ATTO Fluor-488 inhibits K_ir3.2 channels heterologously expressed in Xenopus oocytes.
A continuous current trace recorded at a holding potential of -80 mV. K_ir3.2 currents are downward reflections activated by high K⁺containing solution. While activated, 50 nM and 100 nM of Tertiapin-Q-ATTO Fluor-488 (#STT-170-AG) were applied for 2 min (indicated as bars).

References - Scientific background

Jin, W. and Lu, Z. (1998) Biochemistry 37, 13291.
Drici, M.D. et al. (2000) Br. J. Pharmacol. 131, 569.
Kitamura, H. et al. (2000) J. Pharmacol. Exp. Ther. 293, 196.
Peleg, S. et al. (2002) Neuron 33, 87.
Kanjhan, R. et al. (2005) J. Pharmacol. Exp. Ther. 314, 1353.

Scientific background

Tertiapin, the native toxin, was originally isolated from European honey bee Apis mellifera venom. Native and synthetic Tertiapin blocks a range of inward rectifier K⁺ channels (K_ir), in particular ROMK1 (K_ir1.1, IC₅₀= 2 nM) and GIRK (K_ir3 family, IC₅₀ for the K_ir3.1/3.4 heteromer was 8.6 nM) but with no effect on the K_ir2 family member¹. In accordance, it was shown to inhibit acetylcholine induced K⁺ currents in mammalian cardiomyocytes^2,3.

Tertiapin-Q is a derivative of Tertiapin in which Met¹³ is substituted by a Gln residue. However, unlike native Tertiapin, Tertiapin-Q is non-oxidizable and therefore is more stable⁴.

Tertiapin-Q inhibits the above-mentioned channels with similar affinities and also inhibits Ca²-activated large conductance BK-type K⁺ channels in a concentration and voltage-dependent manner⁵.

Target K_ir1.1, K_ir3.2 K⁺channels

Peptide Content: 100%

Lyophilized Powder

Tertiapin-Q-ATTO Fluor-488 (#STT-170-AG) 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.

Certificate of Analysis SDS

For research purposes only, not for human use

Last Update: 13/06/2024

Applications

Citations

Specifications

Scientific Background

Specific Control Product

Isotype controls are primary antibodies that lack specificity to the target, but match the class and type of the primary antibody used in the application. Isotype controls are used as negative controls to help differentiate non-specific background signal from specific antibody signal.
Our Rabbit IgG Isotype Control-ATTO Fluor-488 (#RIC-001-AG), was specifically developed to be used as a negative control in immunohistochemistry and surface staining flow cytometry applications, along our line of rabbit primary antibodies conjugated to the ATTO Fluor-488 fluorophore.

Rabbit IgG Isotype Control-ATTO Fluor-488 (#RIC-001-AG)

Shipping and Ordering Information

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Tertiapin-Q-ATTO Fluor-488

Overview

Applications

Citations

Specifications

Scientific Background

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