Hi, I am trying to express a HisTag recombinant protein using BL21(De3).
I'd normally grow and induce like 100ml bacteria culture, pellet, then lyse in probably 10ml of lysis buffer. Then sonicate and spin down the cells, which gives me like 10ml supernatant containing the crude protein. From there we'd use HisTag purification kit to isolate the target protein.
I believe that is the standard protocol.
Now I am online researching a good HisTag protein purification kit, and found that most of the minipreps are with each column fits only like 750ul clear lysates.
That makes me wonder why would people purify from such small volume of lysates? How much induced culture (in ml) they used then? Is it enough for the eluted protein to be detected in western blotting?