In addition to all the excellent suggestions to perform seeding, you could also 
try sitting drop vapor diffusion, and also different temperatures. Sometimes 
moving from 20 to 4 degrees does the trick.

Diana

**************************************************
Diana R. Tomchick
Professor
Departments of Biophysics and Biochemistry
UT Southwestern Medical Center
5323 Harry Hines Blvd.
Rm. ND10.214A
Dallas, TX 75390-8816
diana.tomch...@utsouthwestern.edu<mailto:diana.tomch...@utsouthwestern.edu>
(214) 645-6383 (phone)
(214) 645-6353 (fax)

On Sep 9, 2019, at 10:22 AM, Nikolas 
<nikolas.ca...@gmail.com<mailto:nikolas.ca...@gmail.com>> wrote:

Dear crystalgrowers,

I am currently working with a protein that appeared to be friendly but turned 
out it was not the case.
I found myself to face -in the scale up- the opposite of the usual problem of 
nucleation (I really love how this topic finds new ways to make fun of me). In 
24-well plates, hanging-drop, for the same condition but in different drops I 
found few big but intergrown crystals and/or a full with microcrystals. 
Sometimes also in the same well, when having more drops.
I already decreased the concentration to less than 4mg/mL, made small 
adjustments in the optimizations - both with apo and ligand samples, used Al's 
oil.

I have read about the "containerless crystallization" but since I cannot obtain 
the sample myself I would like to know if there are any experiences and/or if 
there are suggestions for solving this problem.

Many thanks!

Best regards,
Nikolas

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