<P> Sandra,</P>
<P>My interpretation of Katie's question was that she wanted to go from HRP to Alk Phos using an ABC detection system only to change the color of visualization. I didn't get the impression that this was for double staining, simply change color of chromogen. So, yes I did suggest that she just simply change the chromogen rather than detection systems. It sounded very logical I thought. </P>
<P>Victoria<BR></P>
<P> <B><I>Sandra Livingston <slivings@hscprime.hsc.usf.edu></I></B> wrote: <BR>
<BLOCKQUOTE style="BORDER-LEFT: #1010ff 2px solid; MARGIN-LEFT: 5px; PADDING-LEFT: 5px"><!doctype html public "-//w3c//dtd html 4.0 transitional//en">Victoria, I am wondering if you meant to suggest that Catherine try the Vector NovaRed that works with the ABC-HRP system instead of switching to the Vector Red which works with the ABC-Alk-Phos. <BR>
<P>Catherine, unless there is some specific reason for switching to Vector Red (prefer that shade of red? want to do double staining with a different systems?), I agree with Victoria about simply switching to a red chromogen with the HRP system. Unlike AEC, the NovaRed CAN be taken through the alcohols and xylene and mounted with a resinous mounting medium without appreciable loss of signal if the slides are not allowed to sit for any length of time in the alcohols. I also agree that it is best to either re-titrate your reagents (if you feel you must change to Alk Phos) or just run a couple of antibodies that you know work well on known positives with both the ABC-HRP and ABC-Alk Phos at the standard dilutions and compare the staining. <BR>
<P>Victoria Baker wrote: <BR>
<BLOCKQUOTE TYPE="CITE">Hi, <BR>
<P>When switching detection systems it is always safer to check your dilutions again and do a titration run. But instead of switching this, why not get the red substrate that Vector has for Alk-Phos? The kit is like $65.00 and will save more in time an effort on your part. I keep both systems in my lab (HRP & Alk Phos) with red chromogen. We've had good results for the most part. <BR>
<P>Good luck. <BR>
<P>Vikki Baker <BR>
<ADDRESS>American Health Foundation</ADDRESS><BR>
<ADDRESS>Valhalla, New York 10595</ADDRESS><BR>
<P><BR> <B><I>"Bennett, Catherine (Katie)" <cbennett@lrri.org></I></B> wrote: <BR>
<BLOCKQUOTE style="BORDER-LEFT: #1010ff 2px solid; MARGIN-LEFT: 5px; PADDING-LEFT: 5px">Advice needed: <BR>
<P>If I switch an IHC procedure over from an ABC-HRP kit to an ABC-Alk Phos <BR>kit, I shouldn't have to adjust my primary and secondary antibody <BR>concentrations, right? (I want to change cromogen labels from DAB to <BR>Vector-Red.) <BR>
<P>TIA. <BR>
<P>********************************* <BR>Catherine "Katie" Bresee Bennett <BR>Sr. Research Technologist <BR>Lovelace Respiratory Research Institute <BR>Albuquerque, New Mexico <BR> </P></BLOCKQUOTE><BR>
<P><BR><BR>
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