Does anyone have the protocol for apoptosis imaging using Annexin V staining on 96 wells plate?
I try Annexin V from Bioact company and following their protocol (they use Nunc 8-well chamber slides):
1. Apoptosis induction
2. Wash with PBS
3. Rinse in wash buffer
4. BSA blocking
5. Incubate with Annexin V
6. Rinse in wash buffer
7. Fix with PFA 4%
8. Rinse in PBS
9. Rinse in H20
But when I observe the results, the control sample had higher fluorescent signal.
Or does anyone suggest me to use any other dishes or slide which are better for this kind of experiment?
Thank you so much!
Hi,
we've used this protocol in our lab:
http://onlinelibrary.wiley.com/doi/10.1002/0471143030.cb1810s47/abstract
http://link.springer.com/article/10.1007%2Fs00204-015-1536-3
and have been further optimizing it (image analysis/data analysis)
best of luck
Hi,
we used Annexin V binding buffer (10 mM HEPES pH 7.4, 140 mM NaCl, 2.5 mM CaCl2) for our assay in 96-well plates for the staining and washes, not PBS. We images the live cells without PFA fixation. We tried with PBS but got no signal.
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