Jurkat cells were treated with serum starvation for 1 hour
followed by adding serum to the culture medium, and culture
for 4 hours before harvesting the cells. Protein extraction
was prepared using 2D lysis buffer using Applied Biomics
protocols. First, sample was run on 2D DIGE (pH4-7)
and the gel image was scanned using Typhoon 9400 (A).
Second, proteins from the 2D gel were transferred
to the membrane and Western Blot was performed using mouse
monoclonal anti-phosphotyrosine antibody (Clone PY20) and
CF488-labeled goat anti-mouse Ig (Biotium, Hayward, California).
The Western Blot image was scanned using Typhoon 9400 (B)