Hello,
I have an Annexin V-FITC antibody that I use to detect apoptosis on the FACS.
I was wondering if I could use the same antibody to stain for apoptosis on histology slides?
Thankyou!
Katherine - I haven't tried this, but would offer a caveat. Remember that Annexin V 'stains' apoptotic cells by engaging phosphatidyl serine that flips from the inner to the outer leaflet of the cell membrane during apoptosis. As such, during flow cytometry Annexin V is useful because it only has access to the extracellular leaflet -- it cant engage inner leaflet PS and so therefore engages only with outer leaflet-displayed PS. In a histological section, Annexin V can access both leaflets of the membrane because the membrane has been sliced during fixation.
You might want to consider a caspase-based apoptosis stain for histology.
Best,
John
John G Younger, MD
www.akadeum.com
Hi Katherine, I have never used for histology slides, but I did use the annexin-FITC for FACS and confocal microscopy, usually with the same sample. That works very well, cause sometimes, for parasites, we can see that annexin isnt only labelling extracellular PS! You should see some immunohistochemistry protocol.
Katherine - I haven't tried this, but would offer a caveat. Remember that Annexin V 'stains' apoptotic cells by engaging phosphatidyl serine that flips from the inner to the outer leaflet of the cell membrane during apoptosis. As such, during flow cytometry Annexin V is useful because it only has access to the extracellular leaflet -- it cant engage inner leaflet PS and so therefore engages only with outer leaflet-displayed PS. In a histological section, Annexin V can access both leaflets of the membrane because the membrane has been sliced during fixation.
You might want to consider a caspase-based apoptosis stain for histology.
Best,
John
John G Younger, MD
www.akadeum.com
Hello
i agree withe researchers 100% , also look at the IHC handbook will help you
Good Luck
http://www.abcam.com/annexin-v-antibody-ab182646.html
Sorry. I'm a little confused. We use annexin-v to detect PS to identify apoptotic cells. How about anti-annexing v antibody? Does that mean you incubate with annexin-v first and then incubate with anti-annexin antibody?
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