Phospho-JNK1/2/3 (T183 + T183 + T221) Recombinant Rabbit Monoclonal Antibody [ST500]

☑ Cell treatment (CT)

Western blot analysis of Phospho-JNK1/2/3 (T183 + T183 + T221) on different lysates with Rabbit anti-Phospho-JNK1/2/3 (T183 + T183 + T221) antibody (ET1609-42) at 1/1,000 dilution.

Lane 1: Hela cell lysate, untreated (10 µg/Lane)
Lane 2: Hela cell lysate, treated with Anisomycin (10 µg/Lane)
Lane 3: A431 cell lysate, untreated (10 µg/Lane)
Lane 4: A431 cell lysate, treated with UV40 (10 µg/Lane)
Lane 5: 293 cell lysate, untreated (10 µg/Lane)
Lane 6: 293 cell lysate, treated with UV40 (10 µg/Lane)
Lane 7: Hela cell lysate, untreated (10 µg/Lane)
Lane 8: Hela cell lysate, treated with UV40 (10 µg/Lane)

Predicted band size: 48/53 kDa
Observed band size: 48/53 kDa

Exposure time: 2 minutes;
12% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1609-42) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

☑ Cell treatment (CT)

Western blot analysis of Phospho-JNK1/2/3 (T183 + T183 + T221) on different lysates with Rabbit anti-Phospho-JNK1/2/3 (T183 + T183 + T221) antibody (ET1609-42) at 1/2,000 dilution.

Lane 1: C6 cell lysate
Lane 2: C6 treated with 25µg/mL Anisomycin for 30 minutes cell lysate.

Lysates/proteins at 20 µg/Lane.

Predicted band size: 48/53 kDa
Observed band size: 48/53 kDa

Exposure time: 2 minutes 6 seconds; ECL: K1802;
4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1609-42) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

☑ Cell treatment (CT)

Western blot analysis of Phospho-JNK1/2/3 (T183 + T183 + T221) on different lysates with Rabbit anti-Phospho-JNK1/2/3 (T183 + T183 + T221) antibody (ET1609-42) at 1/2,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: HeLa treated with 25μg/mL Anisomycin for 30 minutes cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: NIH/3T3 treated with 25μg/mL Anisomycin for 30 minutes cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 48/53 kDa
Observed band size: 48/53 kDa

Exposure time: 59 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1609-42) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

ICC staining of Phospho-JNK1/2/3 (T183 + T183 + T221) in Hela cells (green).

Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1609-42, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

ICC staining of Phospho-JNK1/2/3 (T183 + T183 + T221) in NIH/3T3 cells (green).

Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1609-42, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

ICC staining of Phospho-JNK1/2/3 (T183 + T183 + T221) in HUVEC cells (green).

Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1609-42, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-Phospho-JNK1/2/3 (T183 + T183 + T221) antibody (ET1609-42) at 1/400 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1609-42) at 1/400 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemical analysis of paraffin-embedded human endometrium tissue with Rabbit anti-Phospho-JNK1/2/3 (T183 + T183 + T221) antibody (ET1609-42) at 1/400 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1609-42) at 1/400 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemical analysis of paraffin-embedded mouse heart tissue with Rabbit anti-Phospho-JNK1/2/3 (T183 + T183 + T221) antibody (ET1609-42) at 1/400 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1609-42) at 1/400 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-Phospho-JNK1/2/3 (T183 + T183 + T221) antibody (ET1609-42) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1609-42) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-Phospho-JNK1/2/3 (T183 + T183 + T221) antibody (ET1609-42) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1609-42) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Flow cytometric analysis of Phospho-JNK1/2/3 (T183 + T183 + T221) was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1609-42, 1/100) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).

Immunofluorescence analysis of frozen mouse hippocampus tissue labeling Phospho-JNK1/2/3 (T183 + T183 + T221) with Rabbit anti-Phospho-JNK1/2/3 (T183 + T183 + T221) antibody (ET1609-42).

The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (ET1609-42, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner.

Immunofluorescence analysis of frozen mouse cerebral cortex tissue labeling Phospho-JNK1/2/3 (T183 + T183 + T221) with Rabbit anti-Phospho-JNK1/2/3 (T183 + T183 + T221) antibody (ET1609-42).

The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (ET1609-42, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner.