Originally Posted by Mounir
I'm a master's student in a Biology Departmant. I'm facing a trouble in pre-processing blood samples before using them in Western Blot technique. My aim is to examin the expression of Cathpsin-K, i.e. to measure (approx.) the serum level of this enzyme.
So if anybody can provide me with tips, I would be so pleased.
Welcome to the forum! you have posted in a place that few will be able to help, but I can move your post somewhere else and also help you here.
I assume Cathpsin-K is in the serum? not the blood cells in that case...
As you are using Blood, you would need to:
1) centrifuge the samples (you would need a large amount to begin with)
2) isolate the serum by taking only the supernatant (top layer)
3) it would be difficult to blot so you could load a large amount (or precipitate a larger amount of serum into a smaller volume and load that)
4) blot for that protein
is this a good guide or did I miss anything?