The LDH assay measures cytotoxicity. However, it requires culturing cells in media with less than or equal to 1% FBS. Are there any alternatives?
Yes,
1-When cells become apoptotic, they expose Phosphatidylserine (PS). You can stain for PS using Annexin V that is fluorophore conjugated and run the cells by flow cytometry.
Multiple companies as Biolegend, BD Bioscience, and Thermofisher sell Annexin V staining kits.
2-Another less straightforward way is to perform a western blot or flow cytometry for active capases, such as caspase 3.
Let me know if you need more elaboration.
You can also refer to below given review papers which discuss various methods for detection of apoptosis in addition to the assays mentioned by Mohamed El Tanbouly
1. Curr Protoc Pharmacol. 2004 September 1; 0 12: doi:10.1002/0471141755.ph1208s25.
2. International Dental & Medical Journal of Advanced Research (2016), 2, 1–5.
Dear Chen,
1. Use Trypan blue if FACS sorting is not possible for necrotic cells. FACS sorting would show them in necrotic and apoptotic qads.
2. Use of anti-caspases antibodies for apoptosis.
3. Use Almaar blue to sort viable cells.
I hope this helps. # 1 is when you want to do it at low cost.
Best wishes,
AMB
- Badges
- Science topic
- Similar topics
- Biological Science
- Cell Biology
- Culture Cells