[ccp4bb] In vitro protein interaction analysis using Diazirine crosslinkers.

- Protein crystallography

Main steps:

- Protein purification
- Crystallisation

Special:

- Programs for crystallography
- X-ray detectors

Basic tutorials:

   - Chemistry
   - Protein
   - Peptide
   - Amino Acids

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

Previous message:
Subject: Re: Anomalous map creating
From: Randy Read rjr27 {- at -} CAM {- dot -} AC {- dot -} UK
Date: 2009-11-02

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Subject: Re: Anomalous map creating
From: Guenter Fritz guenter {- dot -} fritz {- at -} UNI-KONSTANZ {- dot -} DE
Date: 2009-11-02

Subject: In vitro protein interaction analysis using Diazirine crosslinkers.
From: Rohan Agashe rohan {- dot -} agashe {- at -} GMAIL {- dot -} COM
Date: 2009-11-02

I hoping for some advice regarding the use of sulfo-NHS diazirine
crosslinker (Pierce) for in vitro protein interaction analysis.

Have tried labeling the protein (8.8kDa, 10 to 20uM conc) with 2 primary
amines using 30-40X of crosslinker in PBS (1hour at 4deg). Have tried other
concentration combinations as well, but this ratio yields some usable
material. However majority of the protein crashes out of solution.

Was wondering if anyone could guide me any tips and tricks in optimizing the
yield of the crosslinked protein?

Thanks in advance.

--
Rohan Agashe

CCP4bb navigation

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

Previous message:
Subject: Re: Anomalous map creating
From: Randy Read rjr27 {- at -} CAM {- dot -} AC {- dot -} UK
Date: 2009-11-02

Next message:
Subject: Re: Anomalous map creating
From: Guenter Fritz guenter {- dot -} fritz {- at -} UNI-KONSTANZ {- dot -} DE
Date: 2009-11-02