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Welcome Protein Crystallography Forum - Protein Crystallography Forum
Welcome Protein Crystallography Forum - Discuss research on protein crystallography and crystalization. Protein Crystallography Forum.
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#1 (permalink) 11-06-2007, 12:34 PM
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Welcome Protein Crystallography Forum
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Welcome to Protein Crystallography Forum.
Please feel free to post questions as threads in this forum about protein crystallization methods and protocols.
#2 (permalink) 11-14-2008, 04:02 PM
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Re: Welcome Protein Crystallography Forum
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Dear All
I wanna crystallize a 18 kDa protein with His-tag & HA tag on C-terminal, and a 19 amino acid signal peptide on N-terminal. My protein is a Singel-Domain antibody. I don't know if they effect on its 3D-structure. My protein with condition has activity. I can't decide that I can crystallize it or I must seprate its tags and signal peptide. Would you please help me?
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Mitra
#3 (permalink) 11-14-2008, 04:31 PM
mitrakheirabadi
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Re: Welcome Protein Crystallography Forum
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Dear All
I wanna crystallize a 18 kDa protein with His-tag & HA tag on C-terminal, and a 19 amino acid signal peptide on N-terminal. My protein is a Singel-Domain antibody. I don't know if they effect on its 3D-structure. My protein with condition has activity. I can't decide that I can crystallize it or I must seprate its tags and signal peptide. Would you please help me?
Best
Mitra
#4 (permalink) 11-14-2008, 04:33 PM
mitrakheirabadi
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Is it correct or not? What do you think?
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Dear All
I wanna crystallize a 18 kDa protein with His-tag & HA tag on C-terminal, and a 19 amino acid signal peptide on N-terminal. My protein is a Singel-Domain antibody. I don't know if they effect on its 3D-structure. My protein with condition has activity. I can't decide that I can crystallize it or I must seprate its tags and signal peptide. Would you please help me?
Best
Mitra
#5 (permalink) 03-01-2009, 04:31 PM
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Re: Welcome Protein Crystallography Forum
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I would remove the tag since it may be controversial if you attain a crystal structure with the tag. For example, a reviewer may bring ask, how do you know that the tag is not influencing your structure? If you don't get any hits without the tag then I would look at the issue again.
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#6 (permalink) 03-11-2009, 02:59 PM
miss deane
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with tag or without tag..
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Hi..
You have two choices whether to crystallize your protein with or without the tag. Both of them might work. If you decide to remove it, then totally remove it from your protein. In my opinion, you can crystallize your protein including the tag. But it would incorporated together with your protein structure, thus it would not show the real structure of your protein. Is that your protein including the tag is 18kDa? The size is small. In this case, you have to make sure that you have enough and high protein content..Because small size of protein will hardly crystallize if it have low protein content..
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Last edited by miss deane; 03-11-2009 at 03:01 PM.
#7 (permalink) 03-18-2009, 04:37 PM
ted9999
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Re: Welcome Protein Crystallography Forum
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I am assuming that high enough protein content means high concentration?
The crystallization of a protein has many variables. I have not read of any articles that indicate protein size (kDa) and protein concentration are directly related. Could you please reference this statement or is this just from personal experience?
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