p38 MAPK, Cell Signaling Technology, 9216

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 1:500 as suggested by the manufacturer.

This antibody has not worked for me using 1:500 dilution. Both TNE lysis buffer and Kinexus lysis buffer was tried with no result.

I also used this antibody on murine samples with a result around the 39 kDa mark.
Konc.: 1:500
Exposure: 30 min
lane-load: 50 ug

Detects a band at around 38 kDa after stimulation that is not present in the unstimulated lane. However, the blot was exploratory and p38 MAPK has not previously been described as activated by this stimulation, meaning it could be an unspecific reaction to a different kinase.
The protein load was 20 ug/lane

WB: 1:500