Re: [ccp4bb] Is there any alternative to siliconized glass coverslips for crystallization?

Long back we used Prosil solution (I think, 1% solution) for siliconization. Mathews -Original Message- From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Goldman, Adrian Sent: Thursday, January 31, 2019 8:43 AM To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] Is

[ccp4bb] Very sorry for one more message == RE: [ccp4bb] Faculty positions at Molecular Biophysics Unit, IISc, Bangalore

I fully agree with Pius. I am very sorry other CCP users for sending one more message and filling your mail box for some crazy guys reply. Thanks, Mathews -Original Message- From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Pius Padayatti Sent: Monday, October

[ccp4bb] Regarding your experience with spectrolight 600 from Molecular Dimensions

Hi Friends, Just checking to see whether any of you have experience with Molecular Dimensions SpectroLight 600 Droplet DLS UV/White light imager or similar systems. Thank you very much, Mathews

[ccp4bb] FW: For 24 well Netxal plates (non ccp4 question) == SUMMARY

Hi Brad, See below my attempts on this in 2010. Regards, Mathews -Original Message- From: Mathews, Irimpan I. Sent: Friday, May 21, 2010 8:23 AM To: CCP4BB@JISCMAIL.AC.UK Subject: RE: For 24 well Netxal plates (non ccp4 question) == SUMMARY Dear All, Thank you for the several

Re: [ccp4bb] Disappearing crystals

Hi Christine, Another minor point (mainly for hanging drops) not mentioned in the messages. When you bring the crystallization tray from lower temperature to room temp. there is a possibility for condensation and slowly droplets near the protein drops gets absorbed into the protein drop. If

Re: [ccp4bb] cryo protection

One small point: Just make sure that you are not too off from the contents of the protein solution.  Sometimes protein solution may have a high amount of salt or things like that and we forget to include atleast half of this concentration into the cryo solution. This could easily crack the

Re: [ccp4bb] Bicarb at low pH

Hi Jacob, Once I had an issue with introuding CO2 into crystals and I was able to get it bound by putting a small piece of dry ice into the well solution and keeping the drop under CO2 pressure using nextal trays with screw cap. Please note in this case CO2 didn't get bound by using pressure

Re: [ccp4bb] Crystal Optimization

Dear Jobi, See if you can slowly increase the PEG concentration to 28 - 30% and that will be a good cryo. Since BIS Tris Propane is the buffer, I think, 28% 3350 should work fine. If the crystals crack by going straight to 28% PEG 3350, 0.1M Bis Tris Propane pH:6.5, 0.2M Potassium thiocyanate

Re: [ccp4bb] Removing a tight binding ligand

Hi Katherine, We had a case where we used saturating amounts of NaCl and precipitated the protein to get rid of a very tight binding ligand (Structure, 11, 677-690, 2003; look at the preparation of the apoenzyme section). Regards, Mathews -Original Message- From: CCP4 bulletin board

Re: [ccp4bb] For 24 well Netxal plates (non ccp4 question) == SUMMARY

-Original Message- From: CCP4 bulletin board [mailto:ccp...@jiscmail.ac.uk] On Behalf Of Mathews, Irimpan I. Sent: Tuesday, May 11, 2010 12:41 PM To: CCP4BB@JISCMAIL.AC.UK Subject: [ccp4bb] For 24 well Netxal plates (non ccp4 question) Dear All, Qiagen has stopped selling the 24 well Netxal

[ccp4bb] For 24 well Netxal plates (non ccp4 question)

Dear All, Qiagen has stopped selling the 24 well Netxal trays and changed to 15 well trays (almost same price) that doesn't fit with any screens. Do you know of some source for the 24 well screw cap trays? Thank you very much for your help, Mathews

Re: [ccp4bb] For 24 well Netxal plates (non ccp4 question)

: Tuesday, May 11, 2010 12:44 PM To: Mathews, Irimpan I. Subject: RE: [ccp4bb] For 24 well Netxal plates (non ccp4 question) They were the only source (sigh!) Maybe they will restart making the original product if enough of us protests loudly enough (hopes not high). Thierry -Original Message

Re: [ccp4bb] Small lysozyme crystals protocol

Hi Jacob, Keep regular drops and run a cat whisker through the drops after 4 - 5 hours. You should have small crystals within 6-10 hrs. If they are still large, use higher protein concentration or precipitant. Kind regards Mathews -Original Message- From: CCP4 bulletin board

Re: [ccp4bb] Crystals grown directly on loop?

Dear Pietro, Thank you very much for sharing your thoughts. I am presuming that you were thinking about microcapillary feeding to reduce the evaporation of the film on the loop. Yes, this is a problem esp. with drops with high salt or alcohol. If there is a way to feed the loop using

Re: [ccp4bb] Crystals grown directly on loop?

Dear Jacob, We have been working on this for an year or so. We have a paper partially in the review process. Unfortunately, the referees were not very excited about the idea. However, we are developing some automation to speed up the time consuming process of placing the drops on the loops.

Re: [ccp4bb] rescuing crashing-out protein eluted from Nickel column

Hi Jacob, You could try keeping some inhibitor or other small molecules that could bind to the protein in the test tube. This may help stabilizing the protein as soon as it comes out of the column. One of my friends had to use an inhibitor during the whole purification procedure (luckily

[ccp4bb]

Hi Daniel, 1. You may try Xenon or Kr pressure cell. We have a simple version of these cells at SSRL (see the link below). Rigaku also has slightly more complex version. Let me know if you want to try this and you don't have access to a pressure cell.

[ccp4bb] Summary for Glove boxes for Crystallization

Hi Friends, Here is a summary for my question regarding Glove boxes for crystallization. They are given approximately in the order they received. I have also kept the name and e-mail address for any additional information. Hope some you will find this information helpful for your research.

[ccp4bb] Question about glove boxes for protein crystallization

Dear Friends, Sorry for the non CCP4 question. We are planning to purchase a small glove box to setup crystallization trays under anaerobic conditions. If you have used glove boxes for crystallization, would you please give me some idea? We are thinking of getting the 815 series from Plas-labs