Re: [ccp4bb] anaerobic crystallization

- Protein crystallography

Main steps:

- Protein purification
- Crystallisation

Special:

- Programs for crystallography
- X-ray detectors

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   - Protein
   - Peptide
   - Amino Acids

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CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Devices Suitable to Concentrate Protein Solutions in Liter Scale
From: Xuan Yang pattisyang {- at -} GMAIL {- dot -} COM
Date: 2009-09-03

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Subject: ´ð¸´: Devices Suitable to Concentrate Protein Solutions in Liter Scale
From: mrmar mrmar2009 {- at -} GMAIL {- dot -} COM
Date: 2009-09-03

Subject: Re: anaerobic crystallization
From: Alexander {- dot -} Schiffer {- at -} SANOFI-AVENTIS {- dot -} COM Alexander {- dot -} Schiffer {- at -} SANOFI-AVENTIS {- dot -} COM
Date: 2009-09-03

Dear Chris,

The answer depends on why you want to exclude oxygen for crystallisation
and how many screens you want to do. Did you purify your protein in the
absence of oxygen?
If you only want to set up a few screens for a single construct - I
would suggest you put the screens and the plates in your anaerobic
chamber a week before you want to use them. Then you set up the drops
manually either in linbro plates or in 96 stripwell plates that you can
seal 8 reservoirs at a time. Depending on the size of your anaerobic
chamber you can also put a microscope in there to look at the drops
(otherwise I would suggest you get plexiglass boxes that you can close
tight to take the plates out for inspection).
I have never tried but it should also be posssible to pipett the drops
with an 8-channel pipett with some practice. You could prefill and seal
the plates outside bring them into the chamber let them equilibrate for
a week and then set up the drops.
I personally think that the Quiagen plates are quite large and you
usually don't have too much storage space in the anaerobic chamber.

Best regards,

Alexander

-----Original Message-----
From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of
Christopher Rife
Sent: Wednesday, September 02, 2009 7:34 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] anaerobic crystallization

Hi,

Does anyone have tips or suggestions for getting started with anaerobic
crystallization? Searching through the archives, I was able to find some
discussion about various glove box options
(http://tinyurl.com/ccp4-glovebox), but not about the process itself (we
already have a box).

To simplify things, would it be worthwhile to perform the initial
screens in something like the pre-filled Qiagen screens
(http://tinyurl.com/qiagen-xseal)? Do I need to worry about evaporation
while solutions are being degassed (esp volatiles)? Better/easier to try
microbatch?

Thanks for any input.

Chris
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CCP4bb navigation

CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Devices Suitable to Concentrate Protein Solutions in Liter Scale
From: Xuan Yang pattisyang {- at -} GMAIL {- dot -} COM
Date: 2009-09-03

Next message:
Subject: ´ð¸´: Devices Suitable to Concentrate Protein Solutions in Liter Scale
From: mrmar mrmar2009 {- at -} GMAIL {- dot -} COM
Date: 2009-09-03