Best way to extract proteins from diatoms

[theguythatknowsnothing]

Hey,

 

I've found some ways to extract proteins from diatoms and I'm going to start trying them. They aren't the easiest to work with but they're fascinating. I was wondering if anyone here has worked with them and knows which one is best. Or even if you have any tips.

 

Greetings

[PBatoon]

What methods have you tried so far? You could probably lyse w/ a salt cut and run it through a purification column. But then I realize you're working with extremely small quantities than recombinant expression. So for diatoms I have no idea. What are you trying to purify?

[Greippi]

There's quite LOADS of literature on the topic. I haven't worked with them, but I'd go with whatever the general literature consensus is. Obviously it depends on WHAT protein you're extracting too - so it might be a bit trial and error finding the optimal technique for different proteins.

[Mr Skeptic]

Yeah, and it would help to know whether you're wanting a membrane protein or a cytoplasmic protein.

[CharonY]

The most robust and universal lysis method is mechanical disruption. But there are literally thousands of protocols for further steps. My main question would be what you are interested in looking at and how. E.g. 2D-PAGE requires something else compared to other chromatographic techniques. Also targeted protein analysis is different from proteome analyses and so on.