Just a thought, but would Testis make a good control block as there are a
lot of cells proliferating and also dying there?
Belfast City Hospital
Colleen Forster wrote:
I stain para fixed mouse tissue with Caspase-3 with good results. The
real key to this stain is to have a CONTROL KNOWN TO HAVE APOPTOTIC
CELLS. I have a transgeneic mouse control that I use and the control
stains beautifully. However, even then there are FEW positive cells.
Another suggestion was to use the intestine as the cells in the
fingerlike crypts do change over at a rapid rate. I have not done my own
trials yet. I have collected the tissue.
I am using the Promega polyclonal Caspase-3 at 1:800. I run the slides
to water, steam with a citrate buffer for 30 min followed by a 10 min
cool down, rinse and stain. I leave my primary on overnight at 4
degrees. I follow with detection from SignetLaboratories, a biotinylated
secondary, streptavidin third, develop with DAB. As a rule you will not
see large numbers of positively stained cells. The staining pattern is
U of MN
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