Nile red.

Tony Houthaeve houthaeve at embl-heidelberg.de
Wed Jun 8 13:29:36 EST 1994


In article <Cr1Dq6.6zK at ncifcrf.gov>, pnh at fcs260c2.ncifcrf.gov (Paul N Hengen) writes:
> 
> I'm thinking of detecting proteins using a dye such as Nile Red or SYBR Red
> from Molecular Probes and then slicing the protein band directly from the gel.
> Is it possible to sequence a protein directly from a polyacryamide gel without
> blotting onto PVDF? Do you foresee any problems with the red dye blocking the
> microsequencing? What is the minimum amount of protein needed for sequencing?
> > ***************************************************************************

It is not possible to sequence a protein within a polyacrylamide gel, so you 
first have to elute the protein out of their or blot it onto PVDF (nitrocellulo
se does not work).
Nile red is a good stain, it has hydrophobic properties and in only a few
minutes a high fluorescence in protein bands in gels is seen.
More it is possible to first stain your gel with Nile red and then additionally
blot onto PVDF. Subsequent sequencing can then be performed, Nile red does not
block.
see : Biotechniques vol 16, No 4 (1994) p 621-624 An article from Bermudez, A,
et al.


Good Luck

Tony Houthaeve
EMBL -Protein & Peptide Group





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