Hi everyone,
I have two questions :
1/ Is there another way besides liquid nitrogen to crush animal tissues stored in the freezer ?
2/ To homogenize samples , which buffer solutions would be most suitable or rather how we choose the right solution to use ?
All that, in order to carry out enzymatic assay tests
thank you in advance for your answers.
Unless the tissue has been dried, it will essentially be a block of ice and any homogenizer will have a fairly difficult time homogenizing it - at least before it thaws. Ordinarily I would recommend a bead mill homogenizer such as one of these: https://homogenizers.net/collections/bead-mill-homogenizers ... but know that the samples will thaw during homogenization.
I'll defer on answering the buffer question specifically, and simply say that homogenizers are not sensitive to the buffer used; you can use whatever buffer is most appropriate for your downstream application.
One additional method that we are successfully following in our lab is using a mechanical homogenizer with a Teflon pestle and glass tube mortar. A small chunk of tissue is removed from storage and added to the tube with a small amount of modified RIPA buffer(with multiple detergents) with protease inhibitor and ground finely. The same is being used by me for heart tissue - proteins for western blot applications. You may consider this method if you do not want to use liquid N2.
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