Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Low endotoxin, Azide free) (ab199376)

Clone EPR25A (ab199376) has been successfully conjugated by Abcam. This image was generated using Rabbit IgG, monoclonal [EPR25A] - Isotype Control (PE). Please refer to ab209478 for protocol details.

Immunofluorescent analysis of HeLa (human cervical cancer) cells, fixed with 4% formaldehyde (10 min). The cells were permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab209478 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control) at 1/500 dilution (showing no signal) and ab195884, Rat monoclonal [YOL1/34] to Tubulin Microtubule Marker (Alexa Fluor® 647), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 100% methanol (5min).

Flow cytometric analysis of 4% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling KCTD5 with ab194825 at 1/70 dilution (red) compared with ab199376 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Low endotoxin, Azide free) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody. ab199376 detects no signal in FC.

Note: ab199376 detects no signal in Flow cytometry.

Clone EPR25A (ab199376) has been successfully conjugated by Abcam. This image was generated using Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647). Please refer to ab199093 for protocol details.

Immunofluorescent analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cells, fixed with 4% formaldehyde (10 min). The cells were permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab199093 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control) at 1/500 dilution (showing no signal) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/250 dilution (shown in green). Nuclear DNA was labeled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 100% methanol (5min).

Clone EPR25A (ab199376) has been successfully conjugated by Abcam. This image was generated using Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 488). Please refer to ab199091 for protocol details.

Immunofluorescent analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cells, fixed with 4% formaldehyde (10 min). The cells were permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab199091 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control) at 1/500 dilution (showing no signal) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 594), at 1/250 dilution (pseudocolored in red). Nuclear DNA was labeled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 100% methanol (5min).

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeled with ab199376 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution. Confocal image showing no staining on HeLa cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows:-
-ve control 1: ab199376 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

Immunohistochemistry analysis of rat colon tissue control for testing ab199376 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. An anti-rabbit IgG H&L (HRP) (ab97051) was used as a secondary antibody. Counterstained with hematoxylin. No staining on rat colon tissue was observed.

Immunohistochemistry analysis of mouse colon tissue control for testing ab199376 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. An anti-rabbit IgG H&L (HRP) (ab97051) was used as a secondary antibody. Counterstained with hematoxylin. No staining on mouse colon tissue was observed.

Immunohistochemistry analysis of human colon tissue control for testing ab199376 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. An anti-rabbit IgG H&L (HRP) (ab97051) was used as a secondary antibody. Counterstained with hematoxylin. No staining on human colon tissue was observed.