All lanes : Anti-JunD antibody [EPR17365] (ab181615) at 1/1000 dilution

Lane 1 : 293T (Human epithelial cells from embryonic kidney) whole cell lysate at 20 µg
Lane 2 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate at 20 µg
Lane 3 : Human fetal liver lysate at 10 µg

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

Predicted band size: 35 kDa
Observed band size: 39,42 kDa why is the actual band size different from the predicted?


Exposure time: 3 minutes

5% NFDM/TBST: Blocking and diluting buffer.

Anti-JunD antibody [EPR17365] (ab181615) at 1/1000 dilution + HeLa (Human epithelial cells from cervix adenocarcinoma ) whole cell lysate at 20 µg

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

Predicted band size: 35 kDa
Observed band size: 39,42 kDa why is the actual band size different from the predicted?


Exposure time: 5 seconds

5% NFDM/TBST: Blocking and diluting buffer.

All lanes : Anti-JunD antibody [EPR17365] (ab181615) at 1/1000 dilution

Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 35 kDa
Observed band size: 39,42 kDa why is the actual band size different from the predicted?


Exposure time: 1 minute

5% NFDM/TBST: Blocking and diluting buffer.

All lanes : Anti-JunD antibody [EPR17365] (ab181615) at 1/1000 dilution

Lane 1 : C6 (Rat glial tumor cells) whole cell lysate
Lane 2 : Raw264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus ) whole cell lysate
Lane 3 : PC12 (Rat adrenal gland pheochromocytoma ) whole cell lysate
Lane 4 : NIH 3T3 (Mouse embyro fibroblast cells ) whole cell lysate

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 35 kDa
Observed band size: 39,42 kDa why is the actual band size different from the predicted?


Exposure time: 30 seconds

5% NFDM/TBST: Blocking and diluting buffer.

Immunohistochemical analysis of paraffin-embedded Human mammary gland tissue labeling JunD using ab181615 at 1/1000 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as secondary at 1/500 dilution. Counterstain: Hematoxylin.
Inset image: negative control obtained using PBS instead of ab181615 and secondary antibody only.
Note: Nuclear staining on the epithelial cells of Human mammary gland was observed.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Immunohistochemical analysis of paraffin-embedded Human lung squamous cell carcinoma tissue labeling JunD using ab181615 at 1/1000 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as secondary at 1/500 dilution. Counterstain: Hematoxylin.
Inset image: negative control obtained using PBS instead of ab181615 and secondary antibody only.
Note: Nucleus staining on the cancer cells of lung squamous cell carcinoma was observed.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Immunohistochemical analysis of paraffin-embedded mouse cerebral cortex tissue labeling JunD using ab181615 at 1/1000 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as secondary at 1/500 dilution. Counterstain: Hematoxylin.
Inset image: negative control obtained using PBS instead of ab181615 and secondary antibody only.
Note: Nuclear staining on neurons of the mouse cerebral cortex was observed.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Immunohistochemical analysis of paraffin-embedded Rat cerebral cortex tissue labeling JunD using ab181615 at 1/1000 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as secondary at 1/500 dilution. Counterstain: Hematoxylin.
Inset image: negative control obtained using PBS instead of ab181615 and secondary antibody only.
Note: Nuclear staining on neurons of the rat cerebral cortex was observed.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells, labeling JunD with ab181615 at 1/10000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image shows nuclear staining on the HeLa cell line. The nuclear counter stain 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 1/500 dilution (red).
The negative controls are as follows:
1. ab181615 at 1/10000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.