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

- Protein crystallography

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

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Subject: Offtopic - In vitro protein interaction analysis using Diazirine crosslinkers.
From: Rohan Agashe rohan {- dot -} agashe {- at -} GMAIL {- dot -} COM
Date: 2009-11-04

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From: P Hubbard dienoylcoa {- at -} HOTMAIL {- dot -} COM
Date: 2009-11-04

Subject: Offtopic - In vitro protein interaction analysis using Diazirine crosslinkers.
From: Rohan Agashe rohan {- dot -} agashe {- at -} GMAIL {- dot -} COM
Date: 2009-11-04

Apologies for an off topic question. I was hoping for some advice regarding
the use of sulfo-NHS diazirine crosslinker (Pierce -
http://www.piercenet.com/products/Browse.cfm?fldID=DB013E61-5056-8A76-4EC4-473A6F2468A1)
for in vitro
protein interaction analysis.

Have tried labeling the protein (9kDa, 10 to 20uM conc) with 2 primary
amines using 30-40X excess Sulfo-NHS-Diazirine crosslinker in PBS (1hour at
4deg). Have tried other concentration combination as well, but this ratio
yields some usable material. However, majority of the protein crashes out of
solution.

Was wondering if anyone has any suggestions in optimizing the yield of
crosslinker labelled protein?

Thanks in advance.
regards,

Rohan Agashe

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

Previous message:
Subject: Offtopic - In vitro protein interaction analysis using Diazirine crosslinkers.
From: Rohan Agashe rohan {- dot -} agashe {- at -} GMAIL {- dot -} COM
Date: 2009-11-04

Next message:
Subject: Off topic - nickel magnetic beads
From: P Hubbard dienoylcoa {- at -} HOTMAIL {- dot -} COM
Date: 2009-11-04