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ab69917 |
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All lanes : Anti-Gli3 antibody (ab69838) at 1 µg/ml
Lane 1 : Pancreas (Mouse) Tissue Lysate
Lane 2 : Liver (Mouse) Tissue Lysate
Lane 3 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 4 : TE 671 (Human Rhabdomyosarcoma) Whole Cell Lysate
Lane 5 : RAW 264.7 (Mouse leukaemic monocyte macrophage cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size : 160 kDa
Observed band size : 145 kDa (why is the actual band size different from the predicted?)
Additional bands at : 110 kDa,55 kDa,75 kDa. We are unsure as to the identity of these extra bands.
ICC/IF image of ab69838 stained RAW 246.7 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 (ab69838, 5µg/ml) overnight at +4ºC. 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. This antibody also gave a positive result in 100% methanol fixed (5 min) RAW 246.7 cells at 5µg/ml.
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