Overview
Phe34 - C-terminal amidation
LC-Biotin
- Ostrow, K.L. et al. (2003) Toxicon 42, 263.
- Suchyna, T.M. et al. (2000) J. Gen. Physiol. 115, 583.
- Redaelli, E. et al. (2010) J. Biol. Chem. 285, 4130.
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.
Live cell imaging of GsMTx-4-Biotin in mouse M1 myeloid leukemia cells.(A-B) CellMask™ Actin 1X solution was applied for 30 minutes, resulting in an orange fluorescence to visualize cellular membrane. (C-D) Following this, the same cells underwent incubation with either 2.5 µM of GsMTx-4-Biotin (C) or GsMTx-4 (#STG-100) (D) for 45 minutes at 37ºC, followed by Alexa Streptavidin-647, leading to red fluorescence indicative of mechanosensitive ion channels distribution. (E-F) Live imaging of M1 cells allowed observation of GsMTx-4 distribution among the cells.
Expression of GsMTx-4-Biotin in Rat DRG Frozen Sections.(A-B) Anti-Piezo1 (extracellular) Antibody (#APC-087) was applied for 60 minutes at 37ºC, followed by staining with goat anti-rabbit AlexaFluor-488, resulting in green fluorescence predominantly in DRG cells. (C-D) The same frozen sections underwent incubation with either 2.5 µM of GsMTx-4-Biotin (C) or GsMTx-4 (#STG-100) (D) for 60 minutes at 37ºC. This was followed by staining with Alexa Streptavidin-647, resulting in red fluorescence in DRG cells. (E-F) Subsequently, CellMask™ Actin 1X solution was applied for 20 minutes to visualize the cellular membrane. (G-H) Co-staining of Anti-Piezo1 (extracellular) Antibody and GsMTx-4 in DRG cells is shown. In (G), the distribution of GsMTx-4 in mechanosensitive ion channels is indicated by arrows, whereas (H) does not show this distribution.
Direct flow cytometry of GsMTx-4 in live intact mouse M1 cells.___ M1 cells.
___ M1 cells + 1.5 µM GsMTx-4 (#STG-100) + 2 µg/ml Streptavidin, Alexa Fluor™ 647 conjugate.
___ M1 cells + 2 µg/ml Streptavidin, Alexa Fluor™ 647 conjugate.
___ M1 cells + 1.5 µM GsMTx-4-Biotin (#STG-100-B) + 2 µg/ml Streptavidin, Alexa Fluor™ 647 conjugate.
Alomone Labs GsMTx-4-Biotin inhibits NaV1.7 channel currents heterologously expressed in Xenopus oocytes.A. Representative time course of GsMTx-4-Biotin (#STG-100-B) inhibition of NaV1.7 channels current. Membrane potential was held at -100 mV, current was elicited by a 100 ms voltage step to 0 mV every 10 sec, and significantly inhibited by application of 0.5 μM GsMTx-4-Biotin (green).
B. Superimposed traces of NaV1.7 channel currents in the absence (control) and presence (green) of 0.5 μM GsMTx-4-Biotin (taken from the recording in A).
- Suchyna, T.M. et al. (2000) J. Gen. Physiol. 115, 583.
- Bode, F. et al. (2001) Nature 409, 35.
- Fang, J. and Iwasa, K.H. (2006) Neurosci. Lett. 404, 213.
- Redaelli, E. et al. (2010) J. Biol. Chem. 285, 4130.
- Spassova, M.A. et al. (2006) Proc. Natl. Acad. Sci. U.S.A. 103, 16586.
- Alessandri-Haber, N. et al. (2009) J. Neurosci. 29, 6217.
- Ostrow, K.L. et al. (2003) Toxicon 42, 263.
- Bae, C. et al. (2011) Biochemistry 50, 6295.
GsMTx-4 is a 34 amino acid peptidyl toxin originally isolated from the Grammostola rosea (Chilean rose) tarantula venom and belongs to the huwentoxin-1 family1.
This toxin inhibits different channels and in addition has antimicrobial activity. It blocks cation-selective mechanosensitive ion channels (strech-activated channels, SACs), without having an effect on whole-cell voltage-sensitive currents1. In addition, it inhibits atrial fibrillation2 as well as the membrane motor of outer hair cells3 at low doses. A medium toxicity on a large spectra of voltage-gated Na+ channels, namely NaV1.1, NaV1.2, NaV1.3, NaV1.4, NaV1.5, NaV1.6 and NaV1.7 was reported. GsMTx-4 also inhibits K+ channels KV11.1 and KV11.2, whereas it does not inhibit K+ channels KV1.1 (IC50 > 85 µM), KV1.4 (IC50 > 85 µM) and KV11.3 (IC50 = 53 µM)4. GsMTx-4 was also found to inhibit both TRPC1 and TRPC6 channels5,6, as well as Piezo1, the mechanosensitive channel7.
Antimicrobial activity is shown against several Gram-positive and Gram-negative bacteria as well8.
GsMTx-4-Biotin (#STG-100-B) is a highly pure, natural, and biologically active conjugated peptide toxin.
Benefits of:
✓ Localization and distribution
✓ Live cell imaging
✓ Single cell detection
✓ Indirect flow cytometry
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