Double IHC question with clarification

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From:C & J Haley <>
To:HistoNet Server <>
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<!doctype html public "-//w3c//dtd html 4.0 transitional//en"> <html> Hello again: <p>Okay, so I didn't give enough information to get enough information. <p>Linda, Patsy, Danielle:      I know, <u>usually</u> if they require the same pretreatment, there's no need  for repeating it.  These guys need to have their own round of EDTA HIER :( . Believe me, I've tried cutting one out,  altering one's time, etc.  I have optimized them separately.  Separately, they look great.   But I need to have the double staining on the same slide.    And that leads me to the next question. <p>Cynthia:  So why would anybody need to put two antibodies that work best alone on the same slide?  The answer is simply:  pesky Pathologist, pretty pictures for a publication, and a nice little challenge.  I should  also mention that one antibody is a nuclear marker and the other is a nice little membranous marker.  Co-expression on the same cell  should make for some nice photos. And to answer your  questions:  yes(substate), yes(times/temps), and no because that shows that you know the many problem parameters of IHC. <p>Diagnostically, most double  immunostains are not practical.  For researchers it can serve many purposes (co-expression, working with material limitations, antibody specificity, etc.).  Also, most  Histotechs (especially those on the Histonet)  like a challenge and a beautifully stained slide. So, I'll continue to "tweak" my procedure and wait for more responses or questions. <br>  <p>Thanks in advance, <p>Jane Haley <br>Barnes-Jewish Hospital <br>St. Louis, Mo. <br>  <br>  <br>  <br> </html>
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