I'm planning to do depolarize neurons. As a positive control, I'm going to use veratrine that can cause voltage‐gated sodium channels to remain open. As a negative control, I need something that block voltage‐gated sodium channels and doesn't allow them to get opened. Is TTX my only choice? do I need to be trained to use it? Does my laboratory need to be special for this purpose?
Thanks in advanced
Hi Mona. You're quite right that TTX is a powerful poison and can be dangerous if not handled carefully, but with appropriate protective measures the risks are manageable.
Essentially, don't touch it, don't get it in your eyes, don't eat it, and don't breathe it in. To achieve this, the equipment needed should be available in any well-equipped laboratory - a locked poisons cupboard for storage, a fume cupboard to handle the material in, and a wash station / eyewash / shower in case of inadvertant exposure. It goes without saying that personal protective equipment should include a lab coat, gloves, and goggles.
A safety data sheet with more details can be found at https://www.caymanchem.com/msdss/14963m.pdf
I'll leave it to others for ideas about alternative agents.
Hope that helps.
TTX can kill you, but so can water. The dose makes the poison.
TTX is toxic to humans in the 10 ug/kg range, with a typical fatal dose being approximately 2-3 mg.
Typically, TTX is provided in 1 mg ampules.
For in vitro usage, one then typically makes 1mM stock aliquots, typically of a 100uL volume, meaning that each aliquot contains 32 micrograms of TTX.
There is no need to find alternatives.
TTX works well and it is very safe to use when handled properly. The working stock solution is dilute enough that it’s not dangerous unless you drink it.
The only tricky part is that it is usually 1mg of powder in a glass ampule. What I do place the tools I need, including a tool to score the glass, the water or buffer solution needed to make a stock solution, the bottle for the stock solution, 2 small Kim Wipes and cellophane tape in the chemical hood where I will open the ampule. I wear gloves, a disposable mask and lab coat as protective barriers.
In the hood, I wrap tape around the neck of the ampule close to where I want to score the glass. After scoring the glass, but before breaking it, I wrap the Kim Wipes around the ampule on either side of the score-line and holding the ampule vertically with the smaller diameter part at the top, I snap the neck and then pour the powder into the stock bottle and add solvent. TTX solution is stable and can be kept refrigerated indefinitely. Mark it with all the necessary hazard symbols.
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