Overview
Lyophilized Powder yes
Origin Synthetic peptide
MW: 3159.6 Da
Purity: >95% (HPLC)
Effective concentration 50-200 nM.
Sequence ALCNCNRIIIPHQCWKKCGKK
Modifications Disulfide bonds between Cys3-Cys14, Cys5-Cys18. Lys21- C-terminal amidation.
Label ATTO-633. ATTO dyes are characterized by strong absorption (high extinction coefficient), high fluorescence quantum yield, and high photo-stability. ATTO-633 has a maximum absorption at 629 nm and a maximum fluorescence at 657 nm. The fluorescence is excited most efficiently in the range 610 to 645 nm. This label is analogous to the well known dye Alexa 647, Alexa 633 and Cy5. 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 (Kir), in particular ROMK1 and GIRK, but with no effect on Kir2 family members1,2. In addition, it was shown to inhibit acetylcholine induced K+ currents in the heart3,4.
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. Ther. 293, 196.
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
Live cell imaging of Tertiapin-Q-ATTO Fluor-633 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 Tertiapin-Q-ATTO Fluor-633 for 30 minutes at 370C, followed by PBSX1 wash, leading to red fluorescence indicative of the distribution of inward rectifier K+ channels (Kir). (C) Live imaging of the differentiated PC-12 cells allowed observation of Tertiapin-Q distribution among the cells.
Unlabeled Tertiapin-Q successfully blocks Kir channels access.(A) Live imaging of the differentiated PC-12 cells allowed observation of Tertiapin-Q-ATTO Fluor-633 (0.5 µM) distribution among the cells. (B) Live imaging of differentiated PC-12 cells demonstrates that Tertiapin-Q (#STT-170, 5 µM) successfully competes with Tertiapin-Q-ATTO Fluor-633 (0.5 µM) for binding sites of Kir channels. (Cells underwent incubation for 15 minutes at 370C).
Direct flow cytometry of Tertiapin-Q in live intact rat PC-12 cells.___ PC-12 cells.
___ PC-12 cells + 50 nM Tertiapin-Q (#STT-170).
___ PC-12 cells + 50 nM Tertiapin-Q-ATTO Fluor-633 (#STT-170-FR).
Alomone Labs Tertiapin-Q-ATTO Fluor-633 inhibits Kir3.2 channel heterologously expressed in Xenopus oocytes.A continuous current trace recorded at a holding potential of -80 mV. Kir3.2 currents are downward reflections activated by high K+ containing solution. While activated, 50 nM and 100 nM of Tertiapin-Q-ATTO Fluor-633 (#STT-170-FR) 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 (Kir), in particular ROMK1 (Kir1.1, IC50 = 2 nM) and GIRK (Kir3 family, IC50 for the Kir3.1/3.4 heteromer was 8.6 nM) but with no effect on the Kir2 family member1. In accordance, it was shown to inhibit acetylcholine induced K+ currents in mammalian cardiomyocytes2,3.
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 Kir1.1, Kir3.2 K+ channels
Peptide Content: 100%
Lyophilized Powder
Tertiapin-Q-ATTO Fluor-633 (#STT-170-FR) is a highly pure, synthetic, and biologically active conjugated peptide toxin.
✓ 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
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