[3dem] Apoferritin for benchmarking

[Raimond NL]

Dear Adam,

Allow us to advertise BfrB, a ferritin from mycobacteria
https://urldefense.proofpoint.com/v2/url?u=http-3A__scripts.iucr.org_cgi-2Dbin_paper-3Fvo5005&d=DwIBaQ&c=-35OiAkTchMrZOngvJPOeA&r=L7-zyQ-04fFCMRqzLIOnx7H0exGZHwIQe_wMPuY600I&m=iM4bVnJ0BkKnOWn-AMW8LOR3FdmEaOiDtkvQzoHJ3iAJnE5S5P6bX37YpDmEWkhY&s=v0B5JqWrTE164UQ_JOGW_DHkKjQYb3PSb_UlLfLWK4I&e= 
We arrived at it after having explored many other ferritins before.
Expression in E-coli, simple ammonium sulfate precipitation, high yield,
decent purity.
High solubility. When used at remarkably high concentrations (eg 50mg/ml)
one gets spectacular densely packed monolayers.
Welcome to request for a plasmid, or to pick up some aliquots in Maastricht.

Best,

Raimond Ravelli
Maastricht University
The Netherlands
++++++++++++++++++++++
Date: Wed, 16 Feb 2022 17:55:12 +0100
From: Adam Schr?fel <adam.schrofel at gmail.com>
To: 3dem at ncmir.ucsd.edu
Subject: [3dem] Apoferritin for benchmarking
Message-ID:
        <CAHx5HtaUnAYGCQuJn=p-A3ONVccRJVvuDy84WyEMMyZGw=g=OA at mail.gmail.com>
Content-Type: text/plain; charset="utf-8"

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
-------------- next part --------------
An HTML attachment was scrubbed...
URL: <http://mail.ncmir.ucsd.edu/pipermail/3dem/attachments/20220217/aef5d452/attachment.html>