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.

• 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.

• IHC-Fr

Lab

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

Immunohistochemical analysis of 4% PFA-fixed 0.2% Triton X-100 permeabilized frozen Rat Brain tissue labeling CD34 with ab81289 at 1/200 (2.4 μg/ml) dilution followed by ab150079 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed at 1/1000 (4 μg/mL). Confocal image showing positive staining on rat brain tissue. The nuclear counterstain was DAPI (Blue). The section was incubated with ab81289 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-Fr

Lab

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

Immunohistochemical analysis of 4% PFA-fixed 0.2% Triton X-100 permeabilized frozen Rat Kidney tissue labeling CD34 with ab81289 at 1/200 (2.4 μg/ml) dilution followed by ab150079 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed at 1/1000 (4 μg/mL). Confocal image showing positive staining on rat kidney tissue. The nuclear counterstain was DAPI (Blue). The section was incubated with ab81289 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-Fr

Lab

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

Immunohistochemical analysis of 4% PFA-fixed 0.2% Triton X-100 permeabilized frozen Mouse Lung tissue labeling CD34 with ab81289 at 1/50 (9.58 μg/ml) dilution followed by ab150079 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed at 1/1000 (4 μg/mL). Confocal image showing positive staining on mouse lung tissue. The nuclear counterstain was DAPI (Blue). The section was incubated with ab81289 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-Fr

Lab

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

Immunohistochemical analysis of 4% PFA-fixed 0.2% Triton X-100 permeabilized frozen Mouse Kidney tissue labeling CD34 with ab81289 at 1/50 (9.58 μg/ml) dilution followed by ab150079 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed at 1/1000 (4 μg/mL). Confocal image showing positive staining on mouse kidney tissue. The nuclear counterstain was DAPI (Blue). The section was incubated with ab81289 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-Fr

Lab

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

Immunohistochemical analysis of 4% PFA-fixed 0.2% Triton X-100 permeabilized frozen Mouse Brain tissue labeling CD34 with ab81289 at 1/50 (9.58 μg/ml) dilution followed by ab150079 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed at 1/1000 (4 μg/mL). Confocal image showing positive staining on mouse brain tissue. The nuclear counterstain was DAPI (Blue). The section was incubated with ab81289 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-Fr

Lab

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

Immunohistochemical analysis of 4% PFA-fixed 0.2% Triton X-100 permeabilized frozen Rat Lung tissue labeling CD34 with ab81289 at 1/200 (2.4 μg/ml) dilution followed by ab150079 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed at 1/1000 (4 μg/mL). Confocal image showing positive staining on rat lung tissue. The nuclear counterstain was DAPI (Blue). The section was incubated with ab81289 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• 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.

• 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)