Donkey Anti-Mouse IgG H&L (Alexa Fluor® 647)

• Flow Cyt

Unknown

Flow Cytometry - Donkey Anti-Mouse IgG H&L (Alexa Fluor® 647) (AB150107)

Overlay histogram showing Jurkat cells stained with ab8090 (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 (ab8090, 0.1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody Donkey anti-mouse IgG H&L (Alexa Fluor^® 647) (ab150107) was used at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 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 solid-state 25mW red diode laser (635nm) and 675/30 bandpass filter.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Donkey Anti-Mouse IgG H&L (Alexa Fluor® 647) (AB150107)

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 / 10% normal goat serum / 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. The secondary antibody (red) was ab150107 Alexa Fluor® 647 donkey anti-mouse IgG (H+L) used at 2μg/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.

• Alexa Fluor®

Unknown

Alexa Fluor® - Donkey Anti-Mouse IgG H&L (Alexa Fluor® 647) (AB150107)

Emmission spectra of the Alexa Fluor^® conjugated secondary antibodies.