Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (AB150061)

ICC/IF image of ab6046 in HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% donkey 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 (ab6046, 1μg/ml) overnight at +4°C. The secondary antibody (green) was ab150061 Alexa Fluor® 488 donkey anti-rabbit IgG (H+L) used at 1μg/ml for 1h. 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.

• Flow Cyt

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Flow Cytometry - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (AB150061)

Overlay histogram showing Jurkat cells stained with ab16669 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal donkey serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab16669, 1/1000 dilution) for 30 min at 22°C. The secondary antibody Donkey anti-rabbit IgG H&L (Alexa Fluor^® 488, pre-adsorbed) (ab150061) was used at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monclonal) (0.1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (AB150061)

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

Unknown

Immunocytochemistry/ Immunofluorescence - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (AB150061)

ICC/IF image of ab6046 stained HeLa 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 (ab6046, 1μg/ml) overnight at +4°C. The secondary antibody (green) was ab150061 Alexa Fluor^® 488 donkey anti-rabbit IgG (H+L) used at 2μg/ml for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.