Donkey Anti-Goat IgG H&L (Alexa Fluor® 594) preadsorbed

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Donkey Anti-Goat IgG H&L (Alexa Fluor® 594) preadsorbed (AB150136)

Composite multiplex immunofluorescence staining of ab302487, ab309493 and ab254351 staining MAP2, Synaptophysin and SV2A in Mouse Primary Neurons DIV14 cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal donkey serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab289874 (shown in yellow), ab309493 (shown in green) and ab254351 (shown in magenta) at 5µg/ml. Cells were then incubated with ab150136 Donkey Anti-Goat IgG H&L (Alexa Fluor® 594) preadsorbed, ab150111 Donkey Anti-Mouse IgG H&L (Alexa Fluor® 647) preadsorbed and ab150061 Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Donkey Anti-Goat IgG H&L (Alexa Fluor® 594) preadsorbed (AB150136)

ICC/IF image of ab7291 stained HeLa cells. The cells were 100% methanol fixed (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then incubated in 1%BSA / 0.3M glycine in 0.1% PBS-Tween for 1h to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab7291, 1μg/ml) overnight at +4°C. ab98800, goat anti-mouse IgG, was then added as a secondary bridging antibody, at 1/250 dilution for 1h. ab150136 Alexa Fluor® 594 donkey anti-goat IgG (H+L) (orange) was then used at 2μg/ml for 1h as a tertiary antibody. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.

The negative control (inset) is a secondary-only assay to demonstrate low non-specific binding of the secondary antibody.

• Alexa Fluor®

Unknown

Alexa Fluor® - Donkey Anti-Goat IgG H&L (Alexa Fluor® 594) preadsorbed (AB150136)