Re: disulphide detection
xiaohui chen (xhchen@biocserver.BIOC.CWRU.Edu)
Mon, 23 Nov 1998 10:39:01 -0500 (EST)
At 11:35 AM 11/23/98 +1000, you wrote:
>Hi everybody.
>We are wondering if anybody knows of a method of detecting disulphide bonds
>(or the lack of) that doesn't involve HPLC or mass spec. We have a 27kDa
>protein with only one disulphide present in the natural state. Is it
>possible to look for a characteristic S-S absorbance by IR or UV? Is there
>a chemical test for disulphides?
>Any suggestions (stupid ones too, they often work best!) would be most welcome.
>Regards
>Michael
>
>
>Michael Jobling
>Dept. of Pathology
>Uni. of Melbourne
>m.jobling@pathology.unimelb.edu.au
>
>
Michael,
Disulfide bond can be easily detected by TNSB assay. Monitor the absorbance
at 412 nm. The reference for TNSB assay is :
Thannhauser, T.W.; Konishi, Y. and Scheraga, H., Analysis for Disulfide
Bonds in Peptides and Proteins, in METHODS IN ENZYMOLOGY (1987), vol. 143.
Good Luck.
Xiaohui Chen, Ph.D.
Department of Biochemistry
School of Medicine
Case Western Reserve University
2109 Adelbert Road
Cleveland, OH 44106
Ph: 216-368-3250
Fax: 216-368-3419
Email: xhchen@biochemistry.cwru.edu