[3dem] molecular weight issue for CryoEM

[Ludovic Renault]

Dear Prakash,

150kDa shouldn't prevent you from trying at least.
Of course you need access to a TEM equipped with a K2 and possibly volta
phase plate.
Before you invest time in the cryo prep, I would advise having a look in
negative stain first, maybe make sure you can get some nice 2d averages or
even a low resolution 3D.
If you don't have a K2/vpp, then collaborate with some lab that has those.
There's also the option to access multi-users cryo-EM facilities. Bob
Glaeser listed a few in commentary of Nat Meth of the year
http://www.nature.com/nmeth/journal/v13/n1/fig_tab/nmeth.3695_T1.html

150 is not easy but it's not impossible, of course it's highly sample
dependent.

Best,
Ludo

On Sat, Apr 23, 2016 at 9:34 PM, prakash shukla <life.prakash at gmail.com>
wrote:

> Dear All,
>
> I have purify a complex of three proteins.They are interacting with each
> other but the molecular weight of protein 150 kDa. I want to do cryoEM for
> complex as for literature molecular weight of protein should be more than
> 300 kDa. how should i proceed for Cryo EM. Suggestions are most welcome.
>
> Regards
>
> Prakash
>
>
>
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