Anti-c-Jun antibody [EP693Y]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-c-Jun antibody [EP693Y] (AB40766)

ab40766 staining c-Jun in wild-type HEK293 cells (top panel) and c-Jun knockout HEK293 cells (bottom panel). The cells were fixed with 100% methanol (5 min) 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 with ab40766 at 1/250 dilution and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at +4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.

Image was taken with a high-content analysis system (Perkin Elmer Operetta CLS™).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-c-Jun antibody [EP693Y] (AB40766)

Intracellular Flow Cytometry analysis of HEK-293 (Human embryonic kidney epithelial cell) cells labeling c-Jun with Purified ab40766 at 1/20 dilution (10μg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluorr^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-c-Jun antibody [EP693Y] (AB40766)

Immunofluorescent staining of HeLa cells
ab40766 at 1/100 dilution

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Jun antibody [EP693Y] (AB40766)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervix carcinoma tissue sections labeling c-Jun with Purified ab40766 at 1 : 500 dilution (0.41 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Jun antibody [EP693Y] (AB40766)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cerebrum tissue sections labeling c-Jun with Purified ab40766 at 1 : 500 dilution (0.41 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-c-Jun antibody [EP693Y] (AB40766)

Immunocytochemistry analysis of NIH/3T3 (Mouse embryonic fibroblast) cells labeling c-Jun with Purified ab40766 at 1 : 50 dilution (4.06 ?g/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1 : 200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1 : 1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Jun antibody [EP693Y] (AB40766)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat liver tissue sections labeling c-Jun with Purified ab40766 at 1 : 500 dilution (0.41 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• WB

Lab

Western blot - Anti-c-Jun antibody [EP693Y] (AB40766)

Lanes 1 - 2 : Merged signal (red and green). Green - ab40766 observed at 35 kDa. Red - loading control, ab18058, observed at 130 kDa.

ab40766 was shown to specifically react with Jun in wild-type HEK-293 cells as signal was lost in Jun knockout cells. Wild-type and Jun knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. ab40766 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-c-Jun antibody [EP693Y] (ab40766) at 1/1000 dilution

Lane 1:

Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 40 µg

Lane 2:

Jun knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 40 µg

Lane 2:

Western blot - Human JUN (c-Jun) knockout HEK-293 cell line (<a href='/en-us/products/cell-lines/human-jun-c-jun-knockout-hek-293-cell-line-ab260862'>ab260862</a>)

Predicted band size: 35 kDa

false

• IP

Lab

Immunoprecipitation - Anti-c-Jun antibody [EP693Y] (AB40766)

Purified ab40766 at 1/20 dilution (1μg) immunoprecipitating c-Jun in NIH/3T3 whole cell lysate.
Lane 1 (input) : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate 10μg
Lane 2 (+) : ab40766 + NIH/3T3 whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab40766 in NIH/3T3 whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 39 kDa

All lanes:

Immunoprecipitation - Anti-c-Jun antibody [EP693Y] (ab40766)

Predicted band size: 35 kDa

false

• WB

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Western blot - Anti-c-Jun antibody [EP693Y] (AB40766)

Blocking Buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-c-Jun antibody [EP693Y] (ab40766) at 1/1000 dilution

Lane 1:

HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate at 15 µg

Lane 2:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 15 µg

Lane 3:

PC-12 (Rat adrenal gland pheochromocytoma ) whole cell lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 35 kDa

Observed band size: 39 kDa

false