Silver Staining Reveals Contaminating Bands in Sample

Paul N Hengen pnh at ncifcrf.gov
Fri Feb 7 15:44:30 EST 1997


Huang Ke-xue (khuang at CHEMVX.CHEM.TAMU.EDU) wrote:

| I am looking for a way to get rid of 2 contaminating bands in one
| of the reagents in SDS Page sample buffer. These bands are visible on
| silver stained gels. I would appreciate hearing from anyone who has had a
| similar experience and has solved this problem. Email replies can be sent
| to klnelson at uvic.ca

: Are those bands in the 60 - 66kDa range?  If yes, then you have just
: re-discovered a famous beta-mercaptoethanol effect, so far,
: unexplainable in its chemistry. 
:
: Your sample buffer or your BME or both are probably to old. Therefore I
: would recommend to prepare fresh buffer. When BME oxidizes it gets
: yellow and has to be discarded.

> I think Dr. Paul wrote a summary about this problem in TIBS (1996). You
> better check the references and do some real work by yourself.

The best explanation I've heard is that the bands come from keratin due
to sloughed human skin cells which make their way into 2-ME and buffer
components. You have to clean and filter everything.

@article{Hengen1996Maytibs,
author = "P. N. Hengen",
title = "Methods and reagents - Eliminating banding
artifacts from {SDS--PAGE}",
journal = "Trends in Biochemical Sciences",
volume = "21",
number = "5",
pages = "191-193",
month = "may",
year = "1996"}

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