This antibody is supposed to detect full-length p55/53, intermediate cleaved p43/41 and complete and active p18. We detect most of the time only p55/53 and p43/41, sometimes at strong apoptotic signals also p18. The antibody produces a lot of unspecific bands, therefore we wash with 0.1% Tween instead of 0.05% Tween and then just use 1:10000 anti-mouse instead of 1:6000, that helps a lot. However especially p43/41 can be weak and not always detectable. Also the antibody tends to precipitate and leave dots, so it might be good to filter it before use. Overall not the best Caspase-8 antibody, but better than e.g. ab32125 which did not work for me.
Use 1:1000 in TBS Tween with 5% BSA, reuse 2-3 times, can be refreshed after that couple of times. Used for cell lines (LNCaP, PC3)
On July 13, 2016 Paula Szalai, University of Oslo wrote:
Caspase-8 (1C12) Cell Signaling Technology #9746 mouse
This antibody is supposed to detect full-length p55/53, intermediate cleaved p43/41 and complete and active p18. We detect most of the time only p55/53 and p43/41, sometimes at strong apoptotic signals also p18. The antibody produces a lot of unspecific bands, therefore we wash with 0.1% Tween instead of 0.05% Tween and then just use 1:10000 anti-mouse instead of 1:6000, that helps a lot. However especially p43/41 can be weak and not always detectable. Also the antibody tends to precipitate and leave dots, so it might be good to filter it before use. Overall not the best Caspase-8 antibody, but better than e.g. ab32125 which did not work for me.
Use 1:1000 in TBS Tween with 5% BSA, reuse 2-3 times, can be refreshed after that couple of times. Used for cell lines (LNCaP, PC3)
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