[Histonet] RE: Vector Red and True Blue

[C.M. van der Loos]

   Hi Ben,

   It's good to read that you're at least successful with the application
   of True Blue since this isn't the most easy chromogen to deal with.
   Furthermore, your choice for this turquoise-red color combination is a
   very good one, because of it's very high contrast.

   Vector Red however, isn't that strong in my hands either. You may try
   to look with a fluorescence microscope as the Vector Red reaction
   product shows a very strong (red) fluorescence upon green light
   excitation (rhodamine-filter pack). Furthermore, I have very good
   results for both light (and fluorescence microscopy) using the new
   Dako Permanent Red kit (K0640). The staining intensity is much
   higher than with Vector Red.

   Chris van der Loos, PhD
   Dept. of Pathology
   Academical Medical Center M2-230
   Meibergdreef 9
   NL-1105 AZ Amsterdam
   The Netherlands

   phone:  +31 20 5665631
   fax:    +31 20 6960389
   e-mail: [1]c.m.vanderloos <@t> amc.uva.nl


   ----- Original Message -----
      From  "Ben" <bjrenquist <@t> ucdavis.edu>
      Date  Wed, 4 May 2005 11:57:08 -0700
        To  <histonet <@t> lists.utsouthwestern.edu>
   Subject  [Histonet] Vector Red and True Blue

   Hello All,

   I  have  been  attempting  some  double  labeling for fos and estrogen
   receptor
   for  about  a month with little luck.  I am using Vector Red (fos) and
   True
   Blue  (ER)  for  my  staining.   I  am  having no problems with the ER
   labeling,
   but  the fos labeling is not that clear.  I don't appear to be getting
   a
   strong reaction product from the Vector Red.  Has anyone else had this
   problem?   How  strong  should the staining be (light pink, dark pink,
   red)?  I
   have been careful to only place in xylene for a few moments before
   coverslipping and therefore don't think fading is my problem.

   I  have been thinking that my antigen retrieval may be the problem.  I
   am
   using triton X to improve antigen retrieval, but was thinking about
   microwaving.   These  are  free  floating  sections.   First  is  this
   possible to
   do with free floati! ng sectio

References

   1. mailto:c.m.vanderloos <@t> amc.uva.nl