Author Topic: Antigen retrieval for TUNEL assay  (Read 7534 times)

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Offline gussy

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Antigen retrieval for TUNEL assay
« on: September 05, 2010, 09:45:40 PM »
Hi all,

Can I use Heat Induce antigen retrieval methods such as using citrate buffer instead of Proteinase K methods in TUNEL assay. My TUNEL kit manual using Proteinase K methods, and I need to buy proteinase K if I follow that manual

Thanks in advance


Antigen retrieval for TUNEL assay
« on: September 05, 2010, 09:45:40 PM »

Offline richard03

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Re: Antigen retrieval for TUNEL assay
« Reply #1 on: September 06, 2010, 12:10:04 PM »
In my experience, the citrate buffer also works for TUNEL staining, but proteinase K seems perform better.

Offline gussy

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Re: Antigen retrieval for TUNEL assay
« Reply #2 on: January 23, 2011, 08:21:46 PM »
Thank you for your kind advice, I have tried and it worked. Now I am performing TUNEL assay on cell culture, I have problem with methyl green counter staining, after 30 seconds of counter staining the stain was very dark, I think my TUNEL is okay because I can see some apoptosis cells, how can I reduce the counter staining color and save my slides?

Any advice will be very appreciate

Regards

Gus

Offline richard03

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Re: Antigen retrieval for TUNEL assay
« Reply #3 on: January 25, 2011, 12:00:42 PM »
Alcohol will remove methyl green quickly so you may want to rinse quickly in 95% alcohol after staining. Refer to this protocol.

http://www.ihcworld.com/_protocols/counterstain_solutions/methyl_green.htm


Re: Antigen retrieval for TUNEL assay
« Reply #3 on: January 25, 2011, 12:00:42 PM »