[ccp4bb] Sonication e.coli parameters

- Protein crystallography

Main steps:

- Protein purification
- Crystallisation

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- Programs for crystallography
- X-ray detectors

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

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From: Dhabaleswar Patra dpatra {- at -} MBU {- dot -} IISC {- dot -} ERNET {- dot -} IN
Date: 2010-04-27

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From: Flip Hoedemaeker flip {- at -} FORMULATRIX {- dot -} COM
Date: 2010-04-27

Subject: Sonication e.coli parameters
From: megha goyal mgbiotec {- at -} GMAIL {- dot -} COM
Date: 2010-04-27

Hi All,

Our protein is expressed as inclusion bodies ibn e.coli W3110 cells. we
harvest the cells supend it in water [no buffer] and perform sonication to
lyse the cells. we lyse the cells till our O.D is decreased to 1/3rd of
original our starting O.D of suspension is about 250. then we centrifuge the
abvoe suspencion at 12000 rpm to collect teh inclusion bodies. but we are
not getting good yield and also inclusion bodies are not that pure. Do we
need to add lysozyme prior to lysis. can anyone suggest a good method to
lyse ecoli cells expressing protein in inclusion bodies.

Thanks in anticipation

meg

CCP4bb navigation

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

Previous message:
Subject:
From: Dhabaleswar Patra dpatra {- at -} MBU {- dot -} IISC {- dot -} ERNET {- dot -} IN
Date: 2010-04-27

Next message:
Subject: Re: X-Ray films
From: Flip Hoedemaeker flip {- at -} FORMULATRIX {- dot -} COM
Date: 2010-04-27