Actinomycin D to 7AAD?

From: Vadim Chromiak (gerund@olinet.isf.kiev.ua)
Date: Fri Mar 07 1997 - 21:55:14 EST


Good morning ladies and gentlemen,

I recently found a lot of Actinomycin D  in  my  lab  (sweet
memory of  the Former SU).  I was somewhat disappointed when
realized  that  unlike  7AAD  it  cannot  be  used  for  DNA
staining. What a pity! I have several grams of that stuff.

Can anybody tell me whether I can use my Actinomycin in some
way,  in relation to flow?  May I use  it,  e.g.,  to  block
Pyronin  Y  binding  to  DNA for measurement of RNA specific
fluorescence? Or  is  it  possible   to   make   7AAD   from
Actinomycin using relatively inexpensive chemicals?  Does it
worth of doing? I suspect that home-made dye may contain too
much  unmodified Actinomycin,  which may reduce DNA staining
competing with 7AAD for binding.  Did anybody in  the  panel
face this problem before?

NB. Please,  don't  send  suggestions like
or  "you  may  use  PI  for DNA staining" (I am not entirely
stupid).

"you may purchase
7AAD from Molecular Probes,  Cat.#00000" (alas,  I may not),

Many thanks in advance!

Vadim Chromyak,
Research Center for Radiology,
Kiev, Ukraine.

PS. I want to thank cordially  Nancy  Perlmutter  and  David
Coder  for  sending  me  protocols and reprints regarding my
question about "DNA plus surface staining".



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