Hmmm…not enough information but I am wondering about the EDTA plus Ca 
business….the Ca++ ions are forming a strong chelate with the EDTA, so

depending on your molar ratios either the free Ca ions would be consumed by 
EDTA or the EDTA by Ca …(?) – any variance 

in ratio here might prove troublesome.

 

BR

From: CCP4 bulletin board [mailto:[email protected]] On Behalf Of dusky dew
Sent: Montag, 14. Juli 2014 13:44
To: [email protected]
Subject: [ccp4bb] cannot reproduce crystals

 

Dear all,
I am trying to reproduce some protein crystals. The protein I am getting after 
cutting the his tag is very pure. I am using the reported protein 
concentration. The cofactor and EDTA needs to be added externally. The 
condition has calcium acetate, peg 4k and sodium acetate buffer. Unfortunately 
I am getting oil separation and light ppt. I have no clue what is wrong. Please 
help! 

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