Overview
Alternative Name G1, Alpha-conotoxin GI
Lyophilized Powder yes
Origin Conus geographus (Geography cone) (Nubecula geographus)
Source Synthetic modified peptide
MW: 2065 Da
Form Lyophilized from double distilled water (ddH2O). May contain TFA as a residual counter ion.
Effective concentration 400 nM- 4 µM
Sequence ECCNPACGRHYSC-NH2
Modifications Cys13 - C-terminal amidation
ATTO-647N
Disulfide bonds between: Cys2-Cys7, Cys3-Cys13
ATTO-647N
Disulfide bonds between: Cys2-Cys7, Cys3-Cys13
Label ATTO-647N. ATTO dyes are characterized by strong absorption (high extinction coefficient), high fluorescence quantum yield, and high photo-stability. Maximum absorption 646 nm; maximum fluorescence 664 nm. The fluorescence is excited most efficiently in the range 625 - 660 nm. A suitable excitation source is the 647 nm line of the Krypton-Ion laser or a diode-laser emitting at 650 nm. It can be used in flow cytometry (FACS) using the red (637 nm) laser. The extent of labeling is one molecule of dye per one α-Conotoxin GI molecule.
Structure
Activity α-Conotoxin GI inhibits α/δ site on mouse muscle-derived BC3H-1 nicotinic acetylcholine receptors (nAChR) expressed on postsynaptic membranes. Other site (α/γ site) on nicotinic receptors from Torpedo californica electric organ is also inhibited by α-Conotoxin GI1,2.
References-Activity
- Groebe, D.R. et al. (1995) Mol. Pharmacol. 48, 105.
- Groebe, D.R. et al. (1997) Biochemistry 36, 6469.
Accession number X5I9Y2
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 Centrifuge the vial before adding solvent (10,000 x g for 5 minutes) to spin down all the powder to the bottom of the vial. The lyophilized product 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.
The product is lyophilized in 0.5 ml conical vial.
The product is soluble in pure water to high-micromolar concentrations (5 µM - 1 mM). For long-term storage in solution, we recommend preparing 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 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.
The product is lyophilized in 0.5 ml conical vial.
The product is soluble in pure water to high-micromolar concentrations (5 µM - 1 mM). For long-term storage in solution, we recommend preparing 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 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 α-Conotoxin GI-ATTO Fluor-647N in live intact human AC16 cells.(A) CellMask™ Actin 1X solution was applied for 30 minutes at 37°C, resulting in a green fluorescence to visualize cellular membrane. (B) Following this, the same cells underwent incubation with 0.4 µM of α-Conotoxin GI-ATTO Fluor-647N (#STC-500-FRN) for 60 minutes at 37°C, leading to red fluorescence. (C) Live imaging of AC16 cells allowed observation of α-Conotoxin GI distribution among the cells.
Unlabeled α-Conotoxin GI successfully blocks α-Conotoxin GI-ATTO Fluor-647N binding.(A) CellMask™ Actin 1X solution was applied for 30 minutes at 37°C, resulting in a green fluorescence to visualize cellular membrane. (B) Following this, the same cells underwent incubation with 20 µM of α-Conotoxin GI (#STC-500) and 0.4 µM of α-Conotoxin GI-ATTO Fluor-647N (#STC-500-FRN) for 60 minutes at 37°C. (C) Live imaging of AC16 cells demonstrates that α-Conotoxin GI competes with α-Conotoxin GI-ATTO Fluor-647N for binding sites.
α-Conotoxin GI-FITC and α-Conotoxin GI-ATTO Fluor-647N stain in mouse heart, L&R ventricle frozen sections.(A-B) Frozen sections underwent incubation with 0.4 µM α-Conotoxin GI-FITC (#STC-500-F) and 0.4 µM α-Conotoxin GI-ATTO Fluor-647N (#STC-500-FRN) for 60 minutes resulting in a green and red fluorescence, respectively. (C) Following this, Hoechst 33342 (2 µg/ml) was applied for 15 minutes, resulting in blue fluorescence to visualize nuclei. (D) Co-localization of α-Conotoxin GI-FITC and α-Conotoxin GI-ATTO Fluor-647N, appears in yellow.
Direct flow cytometry of α-Conotoxin GI in live intact human AC16 cells.___ AC16 cells.
___ AC16 cells + 0.1 µM α-Conotoxin GI (#STC-500).
___ AC16 cells + 0.1 µM α-Conotoxin GI-ATTO Fluor 647N (#STC-700-FRN).
Alomone Labs α-Conotoxin GI-ATTO647N inhibits muscle (α1/β1/δ/γ) nAChR channels expressed in Xenopus oocytes.A. Time course of α-Conotoxin GI-ATTO647N (#STC-500-FRN) inhibition of α1/β1/δ/γ nAChR channels. Membrane potential was held at –80 mV and oocytes were stimulated by exposure to 200 µM acetylcholine every 50 seconds. Application of 300 nM (green) α-Conotoxin GI-ATTO647N significantly inhibits the currents.
B. uperimposed traces of α1/β1/δ/γ nAChR currents upon application of control and of 300 nM (green) α-Conotoxin GI-ATTO647N (taken from the recording in A).
References - Scientific background
- Gray, W.R. et al. (1981) The J. Biol. Chem. 256, 4734.
- Nishiuchi, Y. and Sakakibara, S. (1982) FEBS LETT. 148, 260.
- Guddat, L.W. et al. (1996) Biochemistry 35, 11329.
- Groebe, D.R. et al. (1995) Mol. Pharmacol. 48, 105.
- Groebe, D.R. et al. (1997) Biochemistry 36, 6469.
Scientific background
α-Conotoxin GI is a 13 amino acid peptidyl toxin isolated from the Conus geographus (Geography cone) venom1,2,3. It belongs to the Conotoxin A superfamily and reversibly blocks α/δ nicotinic ACh channel receptors (nAChR).
α-Conotoxin GI reversibly inhibits the high affinity α/δ site on mouse muscle-derived BC3H-1 receptor, a nAChR expressed on postsynaptic membranes4,5. Other low site (α/γ site) on nicotinic receptors from Torpedo californica electric organ is also inhibited by α-Conotoxin GI5.
Target α1/β1/δ/γ nAChR
Lyophilized Powder
α-Conotoxin GI-ATTO Fluor-647N (#STC-500-FRN) is a highly pure, synthetic, and biologically active conjugated toxin.
Benefits of α-Conotoxin GI-ATTO Fluor-647N:
✓ 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.
For research purposes only, not for human use
Last Update: 29/01/2025
Applications
Citations
Citations
Powered by Bioz
