| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IF-Cell, IHC-P |
| Clonality: | Monoclonal |
| Clone number: | SY0297 |
| Form: | Liquid |
| Storage condition: | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term. |
| Storage buffer: | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 36 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Ser63 of Human c-Jun aa 31-80 / 331. |
| Positive control: | C6 treated with 25μg/mL Anisomycin for 30 minutes cell lysate, A549 treated with 250ng/mL anisomycin for 30 minutes whole cell lysate, NIH/3T3 treated with 250ng/mL anisomycin for 30 minutes whole cell lysate, PC-3M, MCF-7, A549, human colon carcinoma tissue, human endometrial tissue. |
| Subcellular location: | Nucleus. |
| Recommended Dilutions:
WB IF-Cell IHC-P |
1:500-1:2,000 1:50-1:200 1:500 |
| Uniprot #: | SwissProt: P05412 Human | P05627 Mouse | P17325 Rat |
| Alternative names: | Activator protein 1 AP 1 AP1 cJun Enhancer Binding Protein AP1 Jun Activation Domain Binding Protein JUN Jun oncogene JUN protein Jun proto oncogene JUN_HUMAN JUNC Oncogene JUN p39 Proto oncogene c jun Proto oncogene cJun Proto-oncogene c-jun Transcription Factor AP 1 Transcription factor AP-1 Transcription Factor AP1 V jun avian sarcoma virus 17 oncogene homolog V jun sarcoma virus 17 oncogene homolog (avian) V jun sarcoma virus 17 oncogene homolog V-jun avian sarcoma virus 17 oncogene homolog vJun Avian Sarcoma Virus 17 Oncogene Homolog |
|
Fig1:
Western blot analysis of Phospho-c-Jun (S63) on different lysates with Rabbit anti-Phospho-c-Jun (S63) antibody (ET1608-4) at 1/2,000 dilution. Lane 1: C6 cell lysate Lane 2: C6 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 Lane 5: NIH/3T3 treated with 25μg/mL Anisomycin for 30 minutes, then treated with λpp for 1 hour cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 36 kDa Observed band size: 40 kDa Exposure time: 1 minute; 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 (ET1608-4) 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. |
|
Fig2: ICC staining of Phospho-c-Jun (S63) in PC-3M 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 (ET1608-4, 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). |
|
Fig3:
Western blot analysis of Phospho-c-Jun (S63) on different lysates with Rabbit anti-Phospho-c-Jun (S63) antibody (ET1608-4) at 1/500 dilution. Lane 1: A549 whole cell lysate Lane 2: A549 treated with 250ng/mL anisomycin for 30 minutes whole cell lysate Lane 3: NIH/3T3 whole cell lysate Lane 4: NIH/3T3 treated with 250ng/mL anisomycin for 30 minutes whole cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 36 kDa Observed band size: 40 kDa Exposure time: 5 minutes; 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 (ET1608-4) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature. |
|
Fig4: ICC staining of Phospho-c-Jun (S63) in MCF-7 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 (ET1608-4, 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). |
|
Fig5: ICC staining of Phospho-c-Jun (S63) in A549 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 (ET1608-4, 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). |
|
Fig6:
Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-Phospho-c-Jun (S63) antibody (ET1608-4) at 1/500 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 (ET1608-4) at 1/500 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. |
|
Fig7:
Immunohistochemical analysis of paraffin-embedded human endometrial tissue with Rabbit anti-Phospho-c-Jun (S63) antibody (ET1608-4) at 1/500 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 (ET1608-4) at 1/500 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. |