This antibody was tested on lysates from primary cultured hippocampal neurons. The antibody was tested on neurons either lysed with TNE buffer (+ complete and phosstop) or with lysis acquired from Kinexus (full recipe can be found here http://www.kinexus.ca/ourServices/immunoblotting/details/index.html ). While we have had success switching between these lysis buffers for detection of specific phosphorylations, this antibody did not work in any of the setting. We tested using both 50ug or 100ug sample pr well, but no bands were observed in any instance. The antibody was tested at a concentration of 1:1000 as suggested by the manufacturer.
On November 29, 2017 Simon Mølgaard wrote:
This antibody was tested on lysates from primary cultured hippocampal neurons. The antibody was tested on neurons either lysed with TNE buffer (+ complete and phosstop) or with lysis acquired from Kinexus (full recipe can be found here http://www.kinexus.ca/ourServices/immunoblotting/details/index.html ). While we have had success switching between these lysis buffers for detection of specific phosphorylations, this antibody did not work in any of the setting. We tested using both 50ug or 100ug sample pr well, but no bands were observed in any instance. The antibody was tested at a concentration of 1:1000 as suggested by the manufacturer.