Re: [ccp4bb] alternatives to IZIT

- Protein crystallography

Main steps:

- Protein purification
- Crystallisation

Special:

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- X-ray detectors

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

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Subject: Re: alternatives to IZIT
From: Richard Gillilan reg8 {- at -} CORNELL {- dot -} EDU
Date: 2008-06-16

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From: "Eric G {- dot -} Huizinga" e {- dot -} g {- dot -} huizinga {- at -} UU {- dot -} NL
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Subject: Re: alternatives to IZIT
From: Artem Evdokimov artem {- at -} XTALS {- dot -} ORG
Date: 2008-06-16

Mercurochrome (mebromin) is both a derivative AND a stain (tested, true).

It's just like having a pie and eating it at the same time.

Artem

-----Original Message-----
From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of
Richard Gillilan
Sent: Sunday, June 15, 2008 9:36 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] alternatives to IZIT

We've tested eight different dyes carefully on three different
proteins and eventually planned to test a much larger number (I''ve
assembled a large collection of dyes of every color and type). So far
everything has worked! (rhodamine, bromophenol blue, and fluorescein
are my favorites because they are fluorophores). You could probably
pick almost anything out of HJ Conn's book "Biological Stains." I
believe there was some trouble with mixes containing high PEG conc,
but I don't recall the details at the moment.

You might even consider using a dye with high bromine content (like
bromophenol blue) to phase the structure, but that would require much
higher concentrations than are typically necessary in ISIT-like
experiments. We did see some evidence of change in crystal morphology
when the proteins where grown with the dyes as opposed to soaked. Not
sure how significant that effect is. Obviously a dye could
potentially induce conformational or oligomeric changes in some cases.

Richard Gillilan
MacCHESS
Ithaca, NY

On Jun 14, 2008, at 4:51 PM, Mark Del Campo wrote:

> Before I place an order for some Izit, are there some other dyes I
> can use to check if I've got a protein
> crystal?
>
> Thanks,
>
> Mark

CCP4bb navigation

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

Previous message:
Subject: Re: alternatives to IZIT
From: Richard Gillilan reg8 {- at -} CORNELL {- dot -} EDU
Date: 2008-06-16

Next message:
Subject: Five positions at Utrecht University
From: "Eric G {- dot -} Huizinga" e {- dot -} g {- dot -} huizinga {- at -} UU {- dot -} NL
Date: 2008-06-16