Annexin V FAQ Analysis and Solutions

When there are abnormal experimental results with Annexin V, should we discard the data and start over? Of course not!

We need to treat failed experimental data seriously, analyze and summarize the possible reasons for the experiment's failure, so that we can re-experiment with a targeted approach and achieve the desired experimental results.

This article mainly analyzes the common problems with Annexin V through practical cases, so that everyone can have a deeper understanding of Annexin V experiments.

Problem 1:

Late-stage apoptotic cells in the treatment group are divided into two groups, lacking early apoptotic cells.

· Probable Cause:

The cell treatment conditions are too harsh, such as: high drug concentration, excessive use of organic solvents for dissolving drugs, extreme conditions (such as boiling water) to treat cells, causing rapid cell death, and presenting a fixed cell effect similar to that in periodic experiments, lacking the apoptosis process.

· Solution:

Treat the cells gently, such as reducing drug concentration; the amount of organic solvent (DMSO, etc.) should be controlled below 5‰.

Problem 2:

Lack of early apoptotic cells, accompanied by a large number of necrotic or naked nuclei cells.

· Probable Cause:

The drug treatment time is too long, and drug-resistant cells turn to naked nuclei after apoptosis, and drug-resistant cells no longer undergo apoptosis.

· Solution:

Shorten the drug treatment time (to ensure the apoptosis effect, the drug treatment dose can be appropriately increased).

Problem 3:

No positive signal in nuclear staining (PI/7-AAD/DAPI).

Question 4:

Indistinct cell grouping

Question 5:

Fluorescence signal in blank group

Question 6:

Large amount of apoptosis in normal cells

If you have encountered other problems in Annexin V experiments, please leave a message below and we will give you feedback as soon as possible.