Re: FW: Caspase-3

From:Geoff McAuliffe

    I don't know why testis would be a good tissue for apoptosis, lots 
of proliferation but not a  lot of apoptosis. Why would cells be dying 
there, other than the usual replacement of 'wear and tear'? Thymus from 
a young animal might be good. Better yet, a PubMed search for "models of 
 apoptosis". Then you would have a paper in a referred journal to cite 
when people started to aruge with you.


Brennan, Liam wrote:

>Just a thought, but would Testis make a good control block as there are a
>lot of cells proliferating and also dying there?
>Liam Brennan
>Histopathology Dept
>Belfast City Hospital
>Northern Ireland
>Colleen Forster wrote:
>Hi Geoff,
>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 
>Colleen Forster
>U of MN

Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029

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