Hi Mike,
We have just recently used TBP for 2D electrophoresis. I read all of the
precautions multiple times with some concern. I transferred the TBP from
the sure seal bottle using three syringe needles inserted in the top. The
first needle was inserted to release any pressure. The second carried a
regulated stream of dry argon to maintain the headspace dry and air-free.
The third needle (long one) was attached to a syringe to withdraw the
required amount of TBP. I wanted a 1 M solution, did the necessary
calculations, and quickly discovered that TBP is not completely soluble nor
miscible with acetonitrile at that concentration. Isopropanol worked very
well. We have directly compared 2mM TBP with 50 mM dithiothreitol in our
BioRad IPG strips. We notice a slightly altered display of protein features
particularly at the higher mass ranges. Lower mass protein constellations
were much less affected. Streaking artifacts in our already fairly good
gels were not at all improved by TBP and in one gel actually looked a little
worse compared to DTT. Your mileage may vary...
Aldrich provides a good description of their SureSeal transfer protocol
along with a little kit. I encourage you to try it and don't remove the
SureSeal cap entirely. Their system is very good at protecting
air-sensitive reagents.
We have been in and out of our stock solution several times without
incident. We maintain a blanket of argon in the headspace of the container.
Wear safety glasses and gloves. Do your transfers in a fume hood.
Best regards to all,
Steve...
"Michael N. Horst, Ph.D." wrote:
> Hello abrfers-
> Over the last few months, I have been considering the use of
> tributyl phosphine (TBP) after reading some encouraging posts to this
> group, regarding its use as a reducing agent in the IEF dimension of 2-D
> PAGE. However, when the reagent arrived here, I learned it is extremely
> air-sensitive and prone to explosion in air. I would like to ask if any
> of you who use this reagent on a regular basis could inform us about
> your routine methods for handling this reagent, diluting it and storing
> the now opened bottle of concentrated reagent. I thank you for your
> time.
> Cheahs,
> Mike
-- Steven H. Seeholzer, Ph.D. ICR Staff Scientist Fox Chase Cancer Center 7701 Burholme Avenue Philadelphia PA 19111voice: (215) 728-1111 fax: (215) 728-3574 email: sh_seeholzer@fccc.edu
trapper
This archive was generated by hypermail 2b29 : Fri Feb 23 2001 - 13:03:34 EST