Is anyone doing the staining for hypoxia using these kits (Hydroxyprobe-1 or Hydroxyprobe Omni)? I have a researcher (his laboratory personnel is not experienced with immunostaining, and not sure he has done it himself) wanting set up immunostaining on murine lung showing hypoxia using one of these kits. One avoids mouse on mouse staining issues (Omni contains polyclonal rabbit primary) while the other kit has success with using the mouse monoclonal primary (Hydroxyprobe-1). For a lab starting out with immunhistochemical staining, avoiding the mouse on mouse issue may be a good introduction to IHC, but maybe the Omni Rabbit polyclonal kit is not as ideal. They can certainly learn the ropes here.
1. Which kit do you prefer?
2. Fluorescence or chromogenic method? With which fixation NBF or frozen sections fixed with acetone?
3. In their website, they cite a protocol by Raleigh et al, where they routinely use CSA (DAKO) for mouse antibodies for clinical samples. Is anyone using CSA as they did? We would like to avoid this if possible, and even use frozen section for enzyme IHC chromogenic methods.
4. Since mouse lung is the target tissue, we would like to use alkaline phosphatase instead of HRP enzyme method? Any comments? Our lung experts prefer alk phos over HRP due to high background issues with peroxidase methods in the past.
Any comments are welcome
Gayle M. Callis
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