[3dem] Apoferritin for benchmarking

[Christos Savva]

Dear Adam

I attach a protocol that Louise Fairall here at Leicester developed a few
years ago. You could just use the same buffer for SEC of your protein. Good
concentrations range between 2-4 mg/ml depending on how concentrated you
want the particles on your grids. Having a protein source with either the
light or heavy chain will give you a better result.

Best wishes

Christos

On Wed, Feb 16, 2022 at 4:55 PM Adam Schröfel <adam.schrofel at gmail.com>
wrote:

> Hello 3DEMers,
> I am wondering if there is any reliable way to prepare apoferritin
> particles for microscope benchmarking and acquisition tuning.
> I've got a A3660 Apoferritin from the equine spleen from Sigma in 50%
> glycerol which is probably the most common source. Is there any protocol on
> how to process the protein? What buffer type I should use and how to get
> rid of glycerol - dialysis, gel filtration? What is the best concentration
> to use, the grid type, etc.? I read some protocols and papers, but the
> processing relatively differs one from another or the protocol is not
> perfectly clear to me.
> Thanks for any suggestions!
> Best regards
>
> Adam
>
> Adam Schröfel
>
> Department of Cell Biology | Biocev
>
> Charles University
>
> Prumyslova 595, 252 50 Vestec
>
> Prague West, Czech Republic
>
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