Gal-T Conditions according to Catherine for big plates
For the Gal-T experiment, beads are further washed 2X in 15 mM Tris pH 7.4 (PMSF+Aprotinin). Then, remove as much fluid as possible using a 30-G needle.
Quantities of antibody to do an IP from a 100 mm plate.
anti-Flag : 4 Ál
anti-Myc : 15 Ál
5172 : 54 Ál
I have checked that it is no need tu use Ram (Rabbit anti-Mouse antibody) to link the beads to these antibodies
Quantity of beads to do an IP from a 100 mm plate:
30-50 Ál (1:1 in PBS + PMSF + Aprotinin)