Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647)

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) (AB150079)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling RFP with ab314225 at 1/2000 (0.252 ug/ml) dilution, followed by ab150079 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) antibody at 1/500 (4 ug/ml) dilution (Green). Confocal image showing positive staining in 293T cells transfected with a RFP expression vector containing a Myc-tag. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). Myc-Tag Mouse mAb (Alexa Fluor® 488) was used to counterstain at 1/100 (0.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150079 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) at 1/500 (4 ug/ml) dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) (AB150079)

ICC/IF image of ab6046 in 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 (ab6046, 2μg/ml) overnight at +4°C. The secondary antibody ab150079 (shown in red) was 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

Supplier Data

Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) (AB150079)

Flow cytometric analysis of HEK-293T (Human embryonic kidney epithelial cell, Left)/ LNCaP (Human prostate carcinoma epithelial cell, Right) cells labelling TMPRSS2 with ab280567 at 1/500 dilution (0.1μg)/ red compared with Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor^® 647, ab150079) at 1/2000 dilution was used as the secondary antibody.

Negative control : HEK-239T (PMID : 20631123).

Gated on viable cells.

• ELISA

Lab

ELISA - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) (AB150079)

Cross-reactivity of the polyclonal secondary antibody ab182016 was tested using a sandwich ELISA approach. The wells were coated with the indicated IgG standards at 1 μg/ml (50 μl/well) and incubated overnight at 4°C, followed by a 5% BSA blocking step for 2h at RT. ab182016 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, ab182016 showed a cross-reactivity of 5-7% towards Human IgG and below 2% towards Mouse IgG, Rat IgG and Chicken IgY.

This data was developed using the unconjugated antibody (ab182016).

• ELISA

Lab

ELISA - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) (AB150079)

Cross-reactivity of Goat anti-Rabbit IgG H&L (ab182016) and Goat anti-Rabbit 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 (50 μl/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.

This data was developed using the unconjugated antibody (ab182016).

• Alexa Fluor®

Unknown

Alexa Fluor® - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) (AB150079)