How ı can do the elisa test of caspase 3 on SH-SY5Y CULTURE. Do someone have the protocol?

Commercially available ELISA kits are available to measure Caspase3. Something like this will work to measure Caspase3 in the cells:

https://www.rndsystems.com/products/human-active-caspase-3-quantikine-elisa-kit_km300

http://www.abcam.com/human-active-caspase-3-ser29-elisa-kit-ab181418.html

You can find the protocols in the product page.

You will need to grow the cells and then lyse them, or break them up, in order to be able to measure the protein levels. Is there something about the procedure that is confusing? There will be a series of steps involved (growing the cells and maintaining them, lysing them, and then the ELISA).

Another easy method of looking at apoptosis is the use of a DAPI and caspase3 stain on your cells, similar to this approach: https://www.moleculardevices.com/en/assets/app-note/dd/img/apoptosis-detection-using-earlytox-caspase-3-7-d-nucview-488-assay-kit-on-imagexpress-micro-systems

Nuclei take on distinct shapes in apoptosis and necrosis and can easily be identified when visualized.

If you have access to a microscope you can easily visualize your cells and count which ones are apoptotic!

You will need to buy antibodies etc. and learn immunocytochemistry, but if you have someone to learn from, it is quite easy.

If you can clarify any of your confusion, please do add to the thread!

All the best,

Elliot

Şeyhmus Bayar

Dear Mr. Glotfelty,

Thank you for your helpfull message. Actualy we buy a caspase-3 kit (sandwich type) from life tein. But I think the protocol of product is not too much clear. I want to read more clear ones. Also I m watching videos about sandwich type elisa. But I didnt saw any video spesific just for caspas-3 elisa. Thank you again. Sincerealy Şeyhmus Bayar

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