Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% tritonX-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling IKK alpha + IKK beta with ab178870 at 1/50 dilution, followed by Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody (ab150077) at 1/200 dilution (green). Nuclear and cytoplasm staining is detected. 

Tubulin is detected with Mouse anti-Tubulin antibody (ab7291) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/400 dilution (red). The nuclear counter stain is DAPI (blue). 

 

The two negative controls were :-

-ve control 1 - ab178870 at 1/50 diltuion followed by Goat anti mouse IgG (Alexa Fluor®594) at 1/400 dilution.
-ve control 2 - Mouse anti-Tubulin antibody (ab7291) at 1/500 dilution followed by Goat anti rabbit IgG (Alexa Fluor®488) at 1/200 dilution.

All lanes : Anti-IKK alpha + IKK beta antibody [EPR16628] (ab178870) at 1/1000 dilution

Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates
Lane 2 : Daudi (Human Burkitt's lymphoma cell line) whole cell lysates
Lane 3 : HL-60 (Human promyelocytic leukemia cells) whole cell lysates

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated antibody at 1/1000 dilution

Predicted band size: 85, 87 kDa
Observed band size: 75,87 kDa why is the actual band size different from the predicted?

Blocking/dilution buffer: 5% NFDM/TBST.

 

ab178870 could recognize 3 isoforms of IKK beta with the MWs of 87kDa, 86kDa and 80kDa, respectively. It could also recognize IKK alpha.

Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling IKK alpha + IKK beta with purified ab178870 at 1/130 dilution(10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

All lanes : Anti-IKK alpha + IKK beta antibody [EPR16628] (ab178870) at 1/1000 dilution

Lane 1 : A431 (Human epidermoid carcinoma) whole cell lysates
Lane 2 : 293T (Human epithelial cells from embryonic kidney) whole cell lysates

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated antibody at 1/1000 dilution

Predicted band size: 85, 87 kDa
Observed band size: 75,87 kDa why is the actual band size different from the predicted?

Blocking/dilution buffer: 5% NFDM/TBST.

ab178870 could recognize 3 isoforms of IKK beta with the MWs of 87kDa, 86kDa and 80kDa, respectively. It could also recognize IKK alpha.

Anti-IKK alpha + IKK beta antibody [EPR16628] (ab178870) at 1/1000 dilution + Human fetal kidney lysates at 20 µg

Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated antibody at 1/1000 dilution

Predicted band size: 85, 87 kDa
Observed band size: 87 kDa why is the actual band size different from the predicted?

Blocking/dilution buffer: 5% NFDM/TBST.

All lanes : Anti-IKK alpha + IKK beta antibody [EPR16628] (ab178870) at 1/1000 dilution

Lane 1 : Mouse brain lysates
Lane 2 : Mouse kidney lysates
Lane 3 : Mouse spleen lysates
Lane 4 : Rat kidney lysates
Lane 5 : C6 (Rat glial tumor cells) whole cell lysates
Lane 6 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysates
Lane 7 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysates

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated antibody at 1/1000 dilution

Predicted band size: 85, 87 kDa
Observed band size: 75,87 kDa why is the actual band size different from the predicted?

Blocking/dilution buffer: 5% NFDM/TBST.

ab178870 could recognize 3 isoforms of IKK beta with the MWs of 87kDa, 86kDa and 80kDa, respectively. It could also recognize IKK alpha.

Immunoprecipitation of Jurkat (Human T cell leukemia cells from peripheral blood) whole cell extracts using ab178870 at 1/40 dilution. Western blot detection was performed using ab178870 at 1/1000 dilution followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated secondary antibody at 1/1000 dilution. The blocking and diluting buffer was 5% NFDM/TBST. 

ab178870 could recognize 3 isoforms of IKK beta with the MWs of 87kDa, 86kDa and 80kDa, respectively. It could also recognize IKK alpha.

 

 

Cross-Immunoprecipitation of Jurkat (Human T cell leukemia cells from peripheral blood) whole cell extracts using ab178870 at 1/40 dilution. Western blot detection was performed using ab32041 (IKK alpha) at 1/1000 dilution followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated secondary antibody at 1/1000 dilution. The blocking and diluting buffer was 5% NFDM/TBST.

This immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% triton-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells is a comparison between ab178870 and a competitor’s leading rabbit polyclonal antibody.

Labeling of IKK alpha + IKK beta with ab178870 is conducted at 1/50 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) secondary antibody (ab150077) at 1/200 dilution.
Labeling of IKK alpha + IKK beta with the competitor antibody is conducted at 1/40 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor ® 488) at 1/200 dilution.

Tubulin is detected with Mouse anti-Tubulin antibody (ab7291) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/400 dilution (red). The nuclear counter stain is DAPI (blue).
The two negative controls were :
-ve control 1 - ab178870 at 1/50 diltuion followed by Goat anti mouse IgG (Alexa Fluor®594) at 1/400 dilution.
-ve control 2 - Mouse anti-Tubulin antibody (ab7291) at 1/500 dilution followed by Goat anti rabbit IgG (Alexa Fluor®488) at 1/200 dilution.

All lanes : Anti-IKK alpha + IKK beta [EPR16628] antibody (ab178870) at 1/1000 dilution, and a competitor's Anti-IKK alpha + IKK beta rabbit polyclonal antibody at 1/200 dilution

Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates
Lane 2 : Daudi (Human Burkitt's lymphoma cell line) whole cell lysates
Lane 3 : HL-60 (Human promyelocytic leukemia cells) whole cell lysates

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated antibody at 1/1000 dilution

Predicted band size: 85, 87 kDa
Observed band size: 75,87 kDa why is the actual band size different from the predicted?

This western blot image is a comparison of ab178870 against a competitor's leading rabbit polyclonal antibody.

Blocking/dilution buffer: 5% NFDM/TBST.

ab178870 could recognize 3 isoforms of IKK beta with the MWs of 87kDa, 86kDa and 80kDa, respectively. It could also recognize IKK alpha.

All lanes : Anti-IKK alpha + IKK beta [EPR16628] antibody (ab178870) at 1/1000 dilution, and a competitor's Anti-IKK alpha + IKK beta rabbit polyclonal antibody at 1/200

Lane 1 : A431 (Human epidermoid carcinoma) whole cell lysates
Lane 2 : 293T (Human epithelial cells from embryonic kidney) whole cell lysates

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated antibody at 1/1000 dilution

Predicted band size: 85, 87 kDa
Observed band size: 75,87 kDa why is the actual band size different from the predicted?

This western blot image is a comparison between ab178870 and a competitor's leading rabbit polyclonal antibody.

Blocking/dilution buffer: 5% NFDM/TBST.

ab178870 could recognize 3 isoforms of IKK beta with the MWs of 87kDa, 86kDa and 80kDa, respectively. It could also recognize IKK alpha.

Anti-IKK alpha + IKK beta [EPR16628] antibody (ab178870) at 1/1000 dilution, and a competitor's Anti-IKK alpha + IKK beta rabbit polyclonal antibody at 1/200 dilution + Human fetal kidney lysates

Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated antibody at 1/1000 dilution

Predicted band size: 85, 87 kDa
Observed band size: 87 kDa why is the actual band size different from the predicted?

This western blot image is a comparison between ab178870 and a competitor's leading rabbit polyclonal antibody.

Blocking/dilution buffer: 5% NFDM/TBST.

All lanes : Anti-IKK alpha + IKK beta [EPR16628] antibody (ab178870) at 1/1000 dilution, and a competitor's Anti-IKK alpha + IKK beta rabbit polyclonal antibody at 1/200 dilution

Lane 1 : Mouse brain lysates
Lane 2 : Mouse kidney lysates
Lane 3 : Mouse spleen lysates
Lane 4 : Rat kidney lysates
Lane 5 : C6 (Rat glial tumor cells) whole cell lysates
Lane 6 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysates
Lane 7 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysates

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated antibody at 1/1000 dilution

Predicted band size: 85, 87 kDa
Observed band size: 75,87 kDa why is the actual band size different from the predicted?

This western blot image is a comparison between ab178870 and a competitor's leading rabbit polyclonal antibody.

Blocking/dilution buffer: 5% NFDM/TBST.

ab178870 could recognize 3 isoforms of IKK beta with the MWs of 87kDa, 86kDa and 80kDa, respectively. It could also recognize IKK alpha.

This immunoprecipation image is a comparison between ab178870 and a competitor's leading rabbit polyclonal antibody.

Immunoprecipitation of Jurkat (Human T cell leukemia cells from peripheral blood) whole cell extracts using ab178870 at 1/40 dilution. Western blot detection was performed using ab178870 at 1/1000 dilution followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated secondary antibody at 1/1000 dilution. The blocking and diluting buffer was 5% NFDM/TBST.

Immunoprecipitation of Jurkat (Human T cell leukemia cells from peripheral blood) whole cell extracts using competitor’s Anti-IKK alpha + IKK beta rabbit polyclonal antibody at 1/12 dilution. Western blot detection was performed using competitor antibody at 1/100 dilution followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated secondary antibody at 1/100 dilution. The blocking and diluting buffer was 5% NFDM/TBST. 

ab178870 could recognize 3 isoforms of IKK beta with the MWs of 87kDa, 86kDa and 80kDa, respectively. It could also recognize IKK alpha.