1. Push a small crystal of Alanine (use a needle etc.) through the oil, positioning it far away from the crystals of protein, as much as possible. It should dissolve in the solution and hopefully it won't harm the protein crystals. Pay attention to which Alanine (or its salt?) you are using because if you're using a salt, it will likely change the pH too much.
2. Set up large-ish drops under oil, use a thin-tip pipettor (tips for 'standard' 1-2ul pipettors can be bought in the 'fine' or 'precision' variety) to add various quantities of Ala, dissolved to saturation in the crystallization matrix (make sure you adjust pH to the original pH of the matrix). Don't rely on pipettor gradations to deliver precise drops (not likely to work under oil and is a crapshoot anyway below 1ul). Instead rely on the visual appearance of drops under oil (i.e. go by size, start small and then increase along a series of drops with crystals) This assumes your apo-crystals are fairly easy to obtain. Artem - Cosmic Cats approve of this message On Mon, Mar 17, 2025 at 8:30 AM srikannathasan velupillai < 0000b52eb5457384-dmarc-requ...@jiscmail.ac.uk> wrote: > Simply transfer the crystals into your stabilization or soaking buffer > using sitting drop or hanging drop plates. I transferred a few drops to > remove the oil, then soaked the ligand, and it worked. You might encounter > some oil droplets, but we were able to manage and successfully collect > data. This method was used for DNA gyrase. Or set up co-crystallisation in > microbatch plate. > Thanks > Kannan > > On Mon, 17 Mar 2025 at 12:14, CENGIZ KAAN FERAH < > 0000a9b927892eef-dmarc-requ...@jiscmail.ac.uk> wrote: > >> Hello everyone, >> >> I'm currently working on a protein and I did solve the apo structure of >> the enzyme. I'm trying to get the ligand bound form of the enzyme. The >> ligand is L-alanine. How should I soak with the ligand? I'm doing >> microbatch under oil with a terasaki plate without any machinery. >> >> Thank you. >> Best Regards. >> >> ------------------------------ >> >> To unsubscribe from the CCP4BB list, click the following link: >> https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 >> > > ------------------------------ > > To unsubscribe from the CCP4BB list, click the following link: > https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 > ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 This message was issued to members of www.jiscmail.ac.uk/CCP4BB, a mailing list hosted by www.jiscmail.ac.uk, terms & conditions are available at https://www.jiscmail.ac.uk/policyandsecurity/
