Recombinant Anti-FAK (phospho Y397) antibody [EP2160Y] - Low endotoxin, Azide free (ab223529)

Unpurified ab81298 staining FAK (phospho Y397) in SK-N-SH (human neuroblastoma cell line) cells treated with anandamide (in water soluble emulsion) (ab120429), by ICC/IF. Increase in FAK (phospho Y397) expression correlates with increased concentration of anandamide (in water soluble emulsion), as described in literature.
The cells were incubated at 37°C for 5 minutes in media containing different concentrations of ab120429 (anandamide (in water soluble emulsion)) in water, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab81298 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue. Membranes are stained in red with WGA.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81298).

Unpurified ab81298 staining FAK (phospho Y397) in SK-N-SH (human neuroblastoma cell line) cells treated with anandamide (ethanol solution) (ab120087), by ICC/IF. Increase in FAK (phospho Y397) expression correlates with increased concentration of anandamide (ethanol solution), as described in literature.
The cells were incubated at 37°C for 10 minutes in media containing different concentrations of ab120087 (anandamide (ethanol solution)) in ethanol, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab81298 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue. Membranes are stained in red with WGA.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81298).

This ICC/IF data was generated using the same anti-FAK (phospho Y397) antibody clone [EP2160Y] in a different buffer formulation (cat# ab81298).

ICC/IF image of unpurified ab81298 stained SKNSH cells. The cells were 100% methanol fixed (5 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 (ab81298, 10µg/ml) overnight at +4°C in PBS containing 1% BSA and 0.1% tween. 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.