Overview
Alternative Name Long neurotoxin 1, α-Bgtx, α-BuTX
Lyophilized Powder yes
Origin Modified natural protein isolated from Bungarus multicinctus (Many-banded krait).
MW: ~9140 Da
Purity: >95% (HPLC)
Form Lyophilized powder.
Effective concentration 1 nM - 3 μM.
Sequence IVCHTTATSPISAVTCPPGENLCYRKMWCDAFCSSRGKVVELGCAATCPSKKPYEEVTCCSTDKCNPHPKQRPG.
Modifications Disulfide bonds between Cys3-Cys23, Cys16-Cys44, Cys29-Cys33, Cys48-Cys59 and Cys60-Cys65.
Label ATTO-633. ATTO dyes are characterized by strong absorption (high extinction coefficient), high fluorescence quantum yield, and high photo-stability. ATTO-633 maximum absorption is 629 nm and maximum fluorescence is at 657 nm. The fluorescence is excited most efficiently in the 610-645 nm range. This label is analogous to the well known dye Alexa 647, Alexa 633 and Cy5. The extent of labeling is 1-3 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 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 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 (50µM-1mM). 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 (50µM-1mM). 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-633 in differentiated PC-12 cells.Neurite outgrowth was induced in PC12 cells through seven days 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 0.5 µM of α-Bungarotoxin-ATTO Fluor-633 for 30 minutes at 37ºC, followed by PBSX1 wash, leading to red fluorescence indicative of the distribution of nicotinic ACh receptors channels. (C) Live imaging of the differentiated PC-12 cells allowed observation of α-Bungarotoxin distribution among the cells.
Expression of α-Bungarotoxin-ATTO Fluor-633 in mouse cortexImmunohistochemical staining of perfusion-fixed frozen mouse brain sections with Anti-Nicotinic Acetylcholine Receptor α7 (CHRNA7) (extracellular) Antibody (#ANC-007), followed by goat-anti-rabbit-AlexaFluor-568, followed by α-Bungarotoxin-ATTO Fluor-633, 0.8 µM. Staining in mouse cortex shows co-staining of apical dendrites (arrow) of pyramidal neurons with α-Bungarotoxin-ATTO Fluor-633 and CHRNA7. Cell nuclei are stained with DAPI (blue).
Alomone Labs α-Bungarotoxin-ATTO Fluor-633 in whole mount staining of mice neuromuscular junction (NMJ)Whole mount staining of mouse neuromuscular junction (NMJ) was stained with the NMJ marker α-Bungarotoxin-ATTO Fluor-633 (#B-100-FR) (purple) at 1 µg/ml concentration. The image was taken using Nikon Epifluorescence microscopy at 60X magnification and is kindly provided by Dr. Eran Perlsson, Dept. of Physiology and Pharmacology, Tel-Aviv University, Tel-Aviv, Israel.
Direct flow cytometry of α-Bungarotoxin in live intact THP-1 monocyte cells___ THP-1 cells.
___ THP-1 cells + 1 µM α-Bungarotoxin (#B-100).
___ THP-1 cells + 1 µM α-Bungarotoxin-ATTO Fluor-633 (#B-100-FR).
Direct flow cytometry of α-Bungarotoxin in live intact rat PC-12 cells___ PC-12 cells.
___ PC-12 cells + 1 µM α-Bungarotoxin (#B-100).
___ PC-12 cells + 1 µM α-Bungarotoxin-ATTO Fluor-633 (#B-100-FR).
Alomone Labs α-Bungarotoxin-ATTO Fluor-633 inhibits muscle nACh receptors heterologously expressed in Xenopus oocytes.A. Time course of muscle nicotinic ACh channel current recording. Membrane potential was held at -80 mV and stimulated every 100 sec with a solution containing 100 µM ACh + 3 μM PNU-120596. 50 nM α-Bungarotoxin-ATTO Fluor-633 (#B-100-FR) was applied (green) for 5 min during the ACh application and inhibited channel current. B. Superimposed examples of muscle nicotinic ACh current in the absence (black) and presence (green) of 50 nM α-Bungarotoxin-ATTO Fluor-633 (taken from the experiment 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
Peptide Content: 100%
Lyophilized Powder
α-Bungarotoxin-ATTO Fluor-633 (#B-100-FR) is a highly pure, natural, and biologically active conjugated peptide toxin.
Benefits of α-Bungarotoxin-ATTO Fluor-633:
✓ Localization and distribution
✓ Clustering and internalization kinetics
✓ Live cell imaging
✓ Single cell detection
✓ Binding kinetics
We gladly take on collaboration projects. Please Contact Us.
For research purposes only, not for human use
Last Update: 25/11/2024
Applications
Citations
Citations
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- Altman, T., et al. (2021) Nat Commun., 12, 6914.
