Hi Leon,

I can recommend the Cryosol kit from Molecular Dimensions:
https://moleculardimensions.com/en/product/MD1-90

https://pmc.ncbi.nlm.nih.gov/articles/PMC5466044/

The use of multi-component solvent mixtures can dramatically boost aqueous 
solubility of otherwise immiscible ligands.
I have had the greatest success with SM1 and SM4.

Key points:

  1.  You need to have a minimum of 40% v/v, and sometimes 50% is better, of 
the solubilising cryo-mixture.
  2.  Crystals are often soluble in well-solution + 40-50% v/v of cryomix.
  3.  So you need to make higher concentration of precipitant such that 
dilution with the cryo-mixture gives you at least the same concentration as in 
the well – going a few % higher is also often advantageous.
  4.  If your crystals grew in a salt based system then I would explore moving 
them into a PEG environment as this will again help with ligand solubiity.
  5.  If your crystals grew from high MW PEG then explore transferring them 
into a higher concentration of lower MW PEG or a PEG smear including low MW 
species as this will again usually increase ligand solubility

Cheers, Charlie.

From: CCP4 bulletin board <[email protected]> On Behalf Of Maria Jose 
Sanchez Barrena
Sent: 13 May 2026 08:00
To: [email protected]
Subject: Re: [ccp4bb] Fragment soaking protein crystals with small molecules

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via the “Report Phishing” button in Outlook.

Hi Leon:We have crystalized several protein-ligand complexes by solubilizing 
the compounds in ethanol. I would try solubility in such solvent.I you need 
DMSO, test how much your protein stands DMSO and you could also add your 
compound (solubilized in 100% DMSO) to your protein.Good luck,Maria ------
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Hi Leon:
We have crystalized several protein-ligand complexes by solubilizing the 
compounds in ethanol. I would try solubility in such solvent.
I you need DMSO, test how much your protein stands DMSO and you could also add 
your compound (solubilized in 100% DMSO) to your protein.
Good luck,
Maria

--------------------
María José Sánchez-Barrena
Department of Crystallography and Structural Biology
Instituto de Química Física “Blas Cabrera”. CSIC.
Calle de Serrano 119. 28006 Madrid
Tel: 0034 915619400 ext. 442058
http://www.xtal.iqf.csic.es/grupo/xmjose/<http://www.xtal.iqfr.csic.es/grupo/xmjose/>
@sanchez_barrena

El 12 may 2026, a las 18:16, Giang, Leon (MU-Student) 
<[email protected]<mailto:[email protected]>>
 escribió:

Hello everyone,

I am looking for advice on my fragment soaking protocol. Recently, I obtained a 
crystal structure of a small molecule within my protein. These compounds are 
insoluble in aqueous solution so I've been preparing aliquots of the compound 
at known concentrations in 100% DMSO.  I aliquot the compounds onto pedestal of 
sitting drop trays then wait for the DMSO to evaporate before adding my 
cryo-protectant solution and pre-grown protein crystals. I am trying to 
reproduce my results with different concentrations of compound but have to wait 
a long time for the DMSO to evaporate. I am looking for advice on how to speed 
up the DMSO evaporation.

Thanks,
Leon

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