ICC/IF image of ab33207 stained MEF1 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab33207, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
All lanes : Anti-EIF2G antibody (ab33207) at 1 µg/mlLane 1 : Mouse Embryonic Fibroblast 1Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate (ab7179)Lysates/proteins at 10 µg per lane.SecondaryIRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/15000 dilutionPerformed under reducing conditions.
All lanes : Anti-EIF2G antibody (ab33207) at 1 µg/mlLane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate (ab7179)Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell LysateLane 3 : Liver (Mouse) Tissue Lysate - normal tissue Lane 4 : Heart (Mouse) Tissue Lysate Lane 5 : Kidney (Mouse) Tissue Lysate Lane 6 : Testis (Mouse) Tissue Lysate - normal tissue Lane 7 : Spinal Cord (Mouse) Tissue LysateLane 8 : Ovary (Mouse) Tissue Lysate - normal tissue Lane 9 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate Lysates/proteins at 10 µg per lane.SecondaryIRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilutionPerformed under reducing conditions.