Practical tips for working with protease inhibitor tablets…
* grind them up first to make it easier to dissolve (I like folding a weigh paper in half, sticking a table in the middle, fold it back in half, then using the spatula to gently grind it up, then pour it into the liquid)
* if you don’t need a whole tablets-worth, make a concentrated stock solution (e.g. dissolve a 50-ml-worth tablet in 2ml of water or 100 mM phosphate buffer, pH 7.0 for 25X) and store aliquots at -20°C for up to 6 months or in the fridge for 1-2 weeks
* always add to your buffers right before you’re ready to use them (for example, if you’re preparing buffers the day before a purification, make the rest of the buffer but hold off on adding the inhibitor)
* if you’re doing something with metal-dependent protein(s) or planning to do a metal affinity chromatography step (e.g. NiNTA for a His-tag purification), make sure to use an EDTA-free one. EDTA is a chelator (metal-biter) so it will steal metals from your proteins (and those of metal-dependent proteases, which is one reason it’s sometimes added) and it will strip the metal off of your column.
here are some more random practical lab tips: [ Ссылка ]
more on chelators: [ Ссылка ] & [ Ссылка ]
more on his-tag purification: [ Ссылка ] & [ Ссылка ] & [ Ссылка ]
more about all sorts of things: #365DaysOfScience All (with topics listed) 👉 [ Ссылка ] or search blog: [ Ссылка ]
