RE: [Histonet] Help; looking for a stain recipe for TTC

From:"Favara, Cynthia (NIH/NIAID)"

You might want to call the author or contact person on the paper. Protocols are usually the  first on the cutting floor. Most people are delighted to have someone pursue their research!
Good luck- c

Cynthia Favara
903 South 4th Street
Hamilton, MT 59840

-----Original Message-----
From: Bernard Ian R SSgt 59 CRES/MSROP [mailto:Ian.Bernard@LACKLAND.AF.MIL]
Sent: Tuesday, October 14, 2003 8:02 AM
To: Histonet (E-mail)
Subject: [Histonet] Help; looking for a stain receipe for TTC
Importance: High

Hello folk or fellow histonetters(in particular in research),
I'm in the process of replicating the histochemical staining method in a protocol article titled " LV-Powered Coronary Sinus Retroperfusion Reduces infarct Size in Acutely Ischemic Pigs."
The made reagents for staining the heart tissue calls for a pre staining is a Phthalol-blue, which we will be substituting with Evans Blue.  I need a concentration and receipe to make this up?   The article does not provide this.  The article states that the " ascending aorta was occluded and phthalo-blue(Evans Blue) was injected into the left ventricle.
Finally the article states: " The left Ventricle was sliced, parallel to the atriopventricular groove, into several 5-10 mm slices and incubated for 30 mins at 37 deg C in TTC or Triphenyltetrazolium Chloride.  There are 2 concentrations by Sigma, which one is to be used here, a 95% or 99.5 %?  Need receipe to make this solution?
What is Trisma HCL and Trisma Base?  I need catalog #'s and source?
The article goes on to state for results:That "the Mycardial tissue containing dehydrogenase enzymes is stained brick red by TTC, but areas of necrotic tissue depleted of these enzymes are not stained and appear as white or pale yellow."
SSgt Bernie

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