Re: [ccp4bb] off topic: Another protein forming gel
-BEGIN PGP SIGNED MESSAGE- Hash: SHA1 Dear Obayed, it is not clear to me whether with 'nothing is working' you mean to concentrate the solution even further or whether you mean that you do not get crystals from 7mg/ml. If you refer to the former, I would proceed with crystallisation trials at 7mg/ml. Tim On 09/21/2011 05:57 AM, Obayed Ullah wrote: Hi all I am working with a human protein (not membrane protein) which is forming gel at higher concentration. I am not able to concentrate the protein even more than 7 mg/ml. It become like jelly which is hard to pipette out. I have gone through all the suggestions which you guys proposed very recently for another member. what i mean that i already have tried different temperature, pH, salt concentration, reducing agents (DTT, TCEF, in different concentration), stability analysis. But unfortunately nothing is working. One think i want to try to add some detergent to it. If you guys have any experience about it then please suggest the name and/or kind of detergent for the protein. Any other suggestion also appreciated which might help. cheers M. Obayed Ullah E.mail: obayed.o...@yahoo.com obayed.o...@gmail.com - -- - -- Dr Tim Gruene Institut fuer anorganische Chemie Tammannstr. 4 D-37077 Goettingen GPG Key ID = A46BEE1A -BEGIN PGP SIGNATURE- Version: GnuPG v1.4.11 (GNU/Linux) Comment: Using GnuPG with Mozilla - http://enigmail.mozdev.org/ iD8DBQFOeaBKUxlJ7aRr7hoRAngZAKD42xoMTzC80OvrOQUwfQ48g4cuRgCg2t5X VVOf9Obqj5eMK5QEwsWbMtQ= =sjqK -END PGP SIGNATURE-
Re: [ccp4bb] off topic: Another protein forming gel
Hi Tim Thanks for your mail. Both are true. I have not got crystals yet. I have tired different truncated constructs as well as concentration for crystallization. Even i set up the tray at 2 mg/ml but still proteins are not mixing well with the drop solutions rather sitting on the middle of the drop. I dont know what to do with that one. Please suggest me. Cheers M. Obayed Ullah E.mail: obayed.o...@yahoo.com obayed.o...@gmail.com - Original Message - From: Tim Gruene t...@shelx.uni-ac.gwdg.de To: Obayed Ullah obayed.o...@yahoo.com Cc: CCP4BB@JISCMAIL.AC.UK Sent: Wednesday, 21 September 2011 6:28 PM Subject: Re: [ccp4bb] off topic: Another protein forming gel -BEGIN PGP SIGNED MESSAGE- Hash: SHA1 Dear Obayed, it is not clear to me whether with 'nothing is working' you mean to concentrate the solution even further or whether you mean that you do not get crystals from 7mg/ml. If you refer to the former, I would proceed with crystallisation trials at 7mg/ml. Tim On 09/21/2011 05:57 AM, Obayed Ullah wrote: Hi all I am working with a human protein (not membrane protein) which is forming gel at higher concentration. I am not able to concentrate the protein even more than 7 mg/ml. It become like jelly which is hard to pipette out. I have gone through all the suggestions which you guys proposed very recently for another member. what i mean that i already have tried different temperature, pH, salt concentration, reducing agents (DTT, TCEF, in different concentration), stability analysis. But unfortunately nothing is working. One think i want to try to add some detergent to it. If you guys have any experience about it then please suggest the name and/or kind of detergent for the protein. Any other suggestion also appreciated which might help. cheers M. Obayed Ullah E.mail: obayed.o...@yahoo.com obayed.o...@gmail.com - -- - -- Dr Tim Gruene Institut fuer anorganische Chemie Tammannstr. 4 D-37077 Goettingen GPG Key ID = A46BEE1A -BEGIN PGP SIGNATURE- Version: GnuPG v1.4.11 (GNU/Linux) Comment: Using GnuPG with Mozilla - http://enigmail.mozdev.org/ iD8DBQFOeaBKUxlJ7aRr7hoRAngZAKD42xoMTzC80OvrOQUwfQ48g4cuRgCg2t5X VVOf9Obqj5eMK5QEwsWbMtQ= =sjqK -END PGP SIGNATURE-
[ccp4bb] off topic: Another protein forming gel
Hi all I am working with a human protein (not membrane protein) which is forming gel at higher concentration. I am not able to concentrate the protein even more than 7 mg/ml. It become like jelly which is hard to pipette out. I have gone through all the suggestions which you guys proposed very recently for another member. what i mean that i already have tried different temperature, pH, salt concentration, reducing agents (DTT, TCEF, in different concentration), stability analysis. But unfortunately nothing is working. One think i want to try to add some detergent to it. If you guys have any experience about it then please suggest the name and/or kind of detergent for the protein. Any other suggestion also appreciated which might help. cheers M. Obayed Ullah E.mail: obayed.o...@yahoo.com obayed.o...@gmail.com
[ccp4bb] off topic: Another protein forming gel
Hi all I am working with a human protein (not membrane protein) which is forming gel at higher concentration. I am not able to concentrate the protein even more than 7 mg/ml. It become like jelly which is hard to pipette out. I have gone through all the suggestions which you guys proposed very recently for another member. what i mean that i already have tried different temperature, pH, salt concentration, reducing agents (DTT, TCEF, in different concentration), stability analysis. But unfortunately nothing is working. One think i want to try to add some detergent to it. If you guys have any experience about it then please suggest the name and/or kind of detergent for the protein. Any other suggestion also appreciated which might help. cheers M. Obayed Ullah E.mail: obayed.o...@yahoo.com obayed.o...@gmail.com
