Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Mouse IgG1 with ab280974 at 1/100 dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The section was incubated with ab280974 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

No staining on the human colon. 

Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells labelling Mouse IgG1 with ab280974 at 1/20 dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) at 1/1000 dilution (Red). The nuclear counterstain was DAPI (Blue).

Confocal image showing no staining in HeLa cells.

Negative control 1: ab280974 at a 1/20 dilution followed by ab150080 at a 1/1000 dilution.

Negative control 2: ab179513 at a 1/200 dilution followed by ab150113 at a 1/1000 dilution.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling Mouse IgG1 with ab280974 at 1/500 dilution (0.1µg) (Red) compared with a Mouse IgG1, kappa monoclonal [MOPC-21] - isotype control ab18443 (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti mouse IgG (Alexa Fluor^® 488, ab150113) at 1/2000 dilution was used as the secondary antibody.

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling Mouse IgG1 with ab280974 at 1/100 dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The section was incubated with ab280974 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

No staining on the mouse kidney. 

Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 cells labelling Mouse IgG1 with ab280974 at 1/20 dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) at 1/1000 dilution (Red). The nuclear counterstain was DAPI (Blue).

Confocal image showing no staining in RAW 264.7 cells.

Negative control 1: ab280974 at a 1/20 dilution followed by ab150080 at a 1/1000 dilution.

Negative control 2: ab179513 at a 1/200 dilution followed by ab1501113 at a 1/1000 dilution.

Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling Mouse IgG1 with ab280974 at 1/100 dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The section was incubated with ab280974 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

No staining on the rat kidney. 

Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized PC-12 cells labelling Mouse IgG1 with ab280974 at 1/20 dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) at 1/1000 dilution (Red). The nuclear counterstain was DAPI (Blue).

Confocal image showing no staining in PC-12 cells.

Negative control 1: ab280974 at a 1/20 dilution followed by ab150080 at a 1/1000 dilution.

Negative control 2: ab179513 at a 1/200 dilution followed by ab150113 at a 1/1000 dilution.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW 264.7 (Human cervix adenocarcinoma epithelial cell, Left) / PC-12 (Rat adrenal gland pheochromocytoma, Right) cells labelling Mouse IgG1 with ab280974 at 1/500 dilution (0.1µg) (Red) compared with a Mouse IgG1, kappa monoclonal [MOPC-21] - isotype control ab18443 (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti mouse IgG (Alexa Fluor^® 488, ab150113) at 1/2000 dilution was used as the secondary antibody.