Goat Anti-Guinea pig IgG H&L (Alexa Fluor® 647) (ab150187)

ICC/IF image of ab7291 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 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 (ab7291, 1µg/ml) overnight at +4°C. The cells were then incubated with ab7075, rabbit anti-mouse HRP conjugated antibody, for 1h at a 1/250 dilution. The cells were then incubated with ab34580, guinea pig anti-HRP antibody, for 1h at a 1/250 dilution. Lastly, the secondary antibody (red) was ab150187 Alexa Fluor® 647 goat anti-guinea pig IgG (H+L) used at 1µg/ml for 1h, as a quaternary antibody. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

ICC image of ab10410 stained SH-SY5Y cells. The cells were fixed with 100% methanol fixed (5 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab10410 (1/200) overnight at +4°C followed by a further incubation at room temperature for 1h with ab150187 Goat Anti-Guinea pig IgG H&L (Alexa Fluor® 647) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.