Overview
Cys26 - C-terminal amidation
ATTO-Fluor 647N
- McDonough, S.I. et al. (1996) J. Neurosci. 16, 2612.
- Herrero, C.J. et al. (1999) Br. J. Pharmacol. 127, 1375.
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.
- Live cell imaging of ω-Conotoxin MVIIC-ATTO Fluor-647N in differentiated PC-12 cells.Neurite outgrowth was induced in PC-12 cells through 8 days exposure to 100 ng/ml Native mouse NGF 2.5S protein (99%) (#N-240). (A-B) CellMask™ Actin 1X solution was applied for 30 minutes, resulting in a green fluorescence to visualize cellular membrane. (C-D) Following this, the same cells underwent incubation with 200 nM of ω-Conotoxin MVIIC-ATTO Fluor-647N (#C-150-FRN), at 37˚C for 5 min leading to red fluorescence indicative of the distribution of CaV2.2 channels. (E-F) Live imaging of differentiated PC-12 cells allowed observation of ω-Conotoxin MVIIC-ATTO Fluor-647N distribution among the cells.
- Unlabeled ω-Conotoxin MVIIC successfully blocks ω-Conotoxin MVIIC-ATTO Fluor-647N binding.Neurite outgrowth was induced in PC-12 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 50 µM of ω-Conotoxin MVIIC (#C-150) and 200 nM of ω-Conotoxin MVIIC-ATTO Fluor-647N (#C-150-FRN), at 37˚C C for 5 min. (C) Live imaging of differentiated PC-12 cells demonstrates that ω-Conotoxin MVIIC successfully competes with ω-Conotoxin MVIIC-ATTO Fluor-647N for binding sites.
- Direct flow cytometry of ω-Conotoxin MVIIC in live intact rat PC-12 cells.___ PC-12 cells.
___ PC-12 cells + 0.1 µM ω-Conotoxin MVIIC (#C-150).
___ PC-12 cells + 0.1 µM ω-Conotoxin MVIIC-ATTO Fluor-647N (#C-150-FRN). - Alomone Labs ω-Conotoxin MVIIC-ATTO Fluor-647N inhibits CaV2.2 channel currents heterologously expressed in Xenopus oocytes.A. Representative time course of ω-Conotoxin MVIIC-ATTO Fluor-647N (#C-150-FRN) inhibition of CaV2.2 channels current. Membrane potential was held at -100 mV, current was elicited by a 100 ms voltage ramp to +60 mV every 10 sec, and inhibited by 1 µM ω-Conotoxin MVIIC-ATTO Fluor-647N (green).
Superimposed traces of CaV2.2 currents after application of control (black) and of 1 µM ω-Conotoxin MVIIC-ATTO Fluor-647N (green), taken from the recording in A.
- McDonough, S.I. et al. (1996) J. Neurosci. 16, 2612.
- Liu, H. et al. (1996) J. Biol. Chem. 23, 13804.
- Stocker, J.W. et al. (1997) J. Neurosci. 17, 3002.
- Herrero, C.J. et al. (1999) Br. J. Pharmacol. 127, 1375.
- Newcomb, R. and Palma, A. (1994) Brain. Res. 638, 95.
- Vega, T. et al. (1995) Eur. J. Pharmacol. 276, 231.
- Hirata, H. et al. (1997) Eur. J. Pharmacol. 321, 217.
ω-Conotoxin MVIIC blocks CaV2.1 (α1A, P/Q-type) and CaV2.2 (α1B, N-type) channels.1 The toxin binds with high affinity to CaV2.1 and with lower affinity to CaV2.2 in rabbit brain.2 However, the block by ω-Conotoxin-MVIIC of N-type channels in DRG neurons developed much faster than the block of P-type currents in Purkinje cells.1 The effect of the toxin is modulated by voltage (i.e. it is more potent for inactivated channels).3
In addition, this toxin was reported to block nicotinic receptors (transiently expressed in Xenopus oocytes) with IC50 of 1.3 µM.4 It was also shown to inhibit K+-induced 3H-GABA release in hippocampus in vivo.5 This effect was with high affinity (50% block, 200 nM). The toxin was used to inhibit synaptic transmission in several peripheral preparations.6,7
ω-Conotoxin MVIIC-ATTO Fluor 647N (#C-150-FRN) is a highly pure, synthetic, and biologically active conjugated toxin.
Benefits of ω-Conotoxin MVIIC-ATTO Fluor 647N:
✓ Localization and distribution
✓ Clustering and internalization kinetics
✓ Live cell imaging
✓ Single cell detection
✓ Binding kinetics
✓ Direct flow cytometry
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