Overview
Alternative Name Long neurotoxin 1, α-Bgtx, α-BuTX
Lyophilized Powder yes
Origin Modified natural protein isolated from Bungarus multicinctus (Many-banded krait).
Source Modified natural protein
MW: ~ 8925 Da
Form Lyophilized Powder
Effective concentration 50 nM – 0.5 µM
Sequence IVCHTTATSPISAVTCPPGENLCYRKMWCDAFCSSRGKVVELGCAATCPSKKPYEEVTCCSTDKCNPHPKQRPG
Modifications Disulfide bonds between: Cys3-Cys23, Cys16-Cys44, Cys29-Cys33, Cys48-Cys and Cys60-Cys65
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 1-2 molecules of dye per molecule α-Bungarotoxin.
Structure
Activity α-Bungarotoxin blocks postsynaptic neuromuscular transmission via competitive inhibition of nicotinic ACh receptors (nAChRs), thereby preventing the depolarizing action on postsynaptic membranes and blocking neuromuscular transmission. Selective for α7 receptors (IC50 value of 1.6 nM) and α3/β4 receptors (IC50 value of >3 μM)1,2. The toxin also blocks GABA(A) receptor subtypes3.
References-Activity
- Wilson, S.P. and Kirshner, N. (1977) J. Neurochem. 28, 687.
- Garcia-Guzman, M. et al. (1995) Eur. J. Neurosci. 7, 647.
- McCann, C.M. et al. (2006) Proc. Natl. Acad. Sci. U.S.A. 103, 5149.
Shipping and storage The product is shipped as a lyophilized powder at room temperature. Upon receipt, the lyophilizate should be stored at -20°C. Protect from moisture. Protect from light and moisture.
Solubility The product is lyophilized in 0.5 ml conical vial.
Centrifuge all products BEFORE adding solvent (10,000 x g for 5 minutes). The preparation of fresh solutions in working buffers before use is recommended. Soluble in pure water to 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 x100-1000 of the final working concentration. Divide the solution into single-use aliquots and store at -20°C. Before use, thaw the relevant vial(s) intended for use and dilute in the desired working buffer. Avoid multiple freeze-thaw cycles to maintain toxin activity.
Centrifuge all products BEFORE adding solvent (10,000 x g for 5 minutes). The preparation of fresh solutions in working buffers before use is recommended. Soluble in pure water to 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 x100-1000 of the final working concentration. Divide the solution into single-use aliquots and store at -20°C. Before use, thaw the relevant vial(s) intended for use and dilute in the desired working buffer. Avoid multiple freeze-thaw cycles to maintain toxin activity.
Storage of solutions Avoid exposure to light. Store the reconstituted solution for the shortest time possible at -20°C. We do not recommend storing the product in working solution for longer than one day. Avoid multiple freeze-thaw cycles.
Our bioassay
Live cell imaging of α-Bungarotoxin-ATTO Fluor-647N in differentiated PC-12 cells.Neurite outgrowth was induced in PC12 cells through 8 days exposure to 100 ng/ml Native mouse NGF 2.5S protein (99%) (#N-240). (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 0.1 µM of α-Bungarotoxin-ATTO Fluor-647N for 60 minutes at 37ºC, followed by PBSX1 wash, leading to red fluorescence indicative of the distribution of nicotinic ACh channels. (C) Live imaging of the differentiated PC-12 cells allowed observation of α-Bungarotoxin distribution among the cells.
Unlabeled α-Bungarotoxin successfully blocks nicotinic ACh channels access.Neurite outgrowth was induced in PC12 cells through 8 days exposure to 100 ng/ml Native mouse NGF 2.5S protein (99%) (#N-240). (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 10 µM of α-Bungarotoxin (#B-100) for 30 minutes at 37ºC, followed by incubation with 0.1 µM of α-Bungarotoxin-ATTO Fluor-647N for 60 minutes at 37ºC, followed by PBSX1 wash. (C) Live imaging of differentiated PC-12 cells demonstrates that α-Bungarotoxin successfully competes with α-Bungarotoxin-ATTO Fluor-647N for binding sites of nicotinic ACh channels.
Direct flow cytometry of α-Bungarotoxin in live intact rat PC-12 cells.___ PC-12 cells.
___ PC-12 cells + 0.1 µM α-Bungarotoxin (#B-100).
___ PC-12 cells + 0.1 µM α-Bungarotoxin-ATTO Fluor-647N (#B-100-FRN).
Direct flow cytometry of α-Bungarotoxin in PC-12-derived extracellular vesicles (EVs).___ PC-12-derived EVs.
___ PC-12-derived EVs + 0.1 µM α-Bungarotoxin (#B-100).
___ PC-12-derived EVs + 0.1 µM α-Bungarotoxin-ATTO Fluor-647N (#B-100-FRN).
Alomone Labs α-Bungarotoxin-ATTO Fluor-647N inhibits α7 nAChR heterologously expressed in Xenopus oocytes.A. Time course of α-Bungarotoxin-ATTO Fluor-647N (#B-100-FRN) action on α7 nAChR currents, elicited every 50 sec by a transient application of 400 µM ACh + 0.3 µM PNU-120596, while membrane potential was held at -80 mV. Application of 10 nM (green) α-Bungarotoxin-ATTO Fluor-647N significantly inhibits the currents. B. Superimposed traces of α7 nAChR currents upon application of control and 10nM (green) α-Bungarotoxin-ATTO Fluor-647N (taken from the recording in A).
References - Scientific background
- Ohta, M. et al. (1987) FEBS Lett. 222, 79.
- Wilson, P.T. et al. (1988) Mol. Pharmacol. 34, 643.
- Wilson, S.P. and Kirshner, N. (1977) J. Neurochem. 28, 687.
- Garcia-Guzman, M. et al. (1995) Eur. J. Neurosci. 7, 647.
- McCann, C.M. et al. (2006) Proc. Natl. Acad. Sci. U.S.A. 103, 5149.
Scientific background
α-Bungarotoxin isoform A31 is a 74 amino acid peptidyl toxin isolated from the venom of the banded krait snake, Bungarus multicinctus1.
α-Bungarotoxin blocks postsynaptic neuromuscular transmission via competitive inhibition of nicotinic ACh receptors (nAChRs) with an IC50 of 3.5 x 10-10 M, thereby preventing the depolarizing action on postsynaptic membranes and blocking neuromuscular transmission2.
The toxin is selective for α7 receptors (IC50 value of 1.6 nM) and α3/β4 receptors (IC50 value of >3 µM)3,4.
α-Bungarotoxin also binds to and blocks a subset of GABAA receptors (GABAARs) that contain the GABAAR β3 subunit. In particular, α-Bungarotoxin blocks GABAARs that contain interfaces between adjacent β3 subunits5.
Target α7, α1/β1/γ/δ nAChR and GABA(A) receptor subtypes
Lyophilized Powder
α-Bungarotoxin-ATTO Fluor-647N (#B-100-FRN) is a highly pure, natural, and biologically active conjugated peptide toxin.
Benefits of α-Bungarotoxin-ATTO Fluor-647N:
✓ Localization and distribution
✓ Live cell imaging
✓ Single cell detection
✓ Direct flow cytometry
✓ Binding kinetics
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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