Goat Anti-Mouse IgG H&L (Alexa Fluor® 405)

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 405) (AB175660)

ICC/IF image of ab7291 stained HeLa cells. The cells were 4% formaldehyde fixed (10 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 (blue) was ab175660 Alexa Fluor^® 405 goat anti-mouse IgG (H+L) used at 1μg/ml for 1h. DRAQ5™ (ab108410) was used to stain the cell nuclei (red) at a concentration of 1.25μM.

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

• sELISA

Lab

Sandwich ELISA - Goat Anti-Mouse IgG H&L (Alexa Fluor® 405) (AB175660)

Cross-reactivity of the polyclonal secondary antibody ab182017 was tested using a sandwich ELISA approach. The wells were coated with the indicated IgG standards at 1 µg/ml (50µl/well) and incubated overnight at 4°C, followed by a 5% BSA blocking step for 2h at RT. ab182017 was then added starting at 1 µg/ml and gradually diluted 1/4 (50 µl/well), followed by incubation for 2h. For the detection Donkey anti-Goat IgG H&L (HRP) (ab6885) was used at 1/10,000 dilution (50 µl/well), followed by incubation for 1h at RT.

For the batch tested, ab182017 showed a cross-reactivity below 2% towards Chicken IgY, 6% towards Human IgG, 7% towards Rabbit IgG and 47% towards Rat IgG.

This data were developed using the unconjugated antibody (ab182017).

• sELISA

Lab

Sandwich ELISA - Goat Anti-Mouse IgG H&L (Alexa Fluor® 405) (AB175660)

Cross-reactivity of Goat anti-Mouse IgG H&L (ab182017) and Goat anti-Mouse IgG H&L obtained from two different vendors was tested using a sandwich ELISA approach. The wells were coated with the indicated IgG standards (Rabbit, Human, Mouse and Rat) at 1 µg/ml (50µl/well) and incubated overnight at 4°C, followed by a 5% BSA blocking step for 2h at RT. Secondary antibodies were then added starting at 1 µg/ml and gradually diluted 1/4 (50 µl/well), followed by incubation for 2h. For the detection Donkey anti-Goat IgG H&L (HRP) (ab6885) was used at 1/10,000 dilution (50 µl/well), followed by incubation for 1h at RT. This data is from a representative dilution.

These data were developed using the unconjugated antibody (ab182017).

• Alexa Fluor®

Unknown

Alexa Fluor® - Goat Anti-Mouse IgG H&L (Alexa Fluor® 405) (AB175660)