Phospho-c-Jun (S63) Recombinant Rabbit Monoclonal Antibody [SY0297]
cat.: ET1608-4
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: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
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 breast carcinoma tissue, human tonsil tissue, 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
1:50-1:500
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
Images
ET1608-4_1.jpg 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.
ET1608-4_2.jpg 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).
ET1608-4_3.jpg 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.
ET1608-4_4.jpg 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).
ET1608-4_5.jpg 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).
ET1608-4_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-Phospho-c-Jun (S63) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-4, 1/50) for 30 minutes 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.
ET1608-4_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Phospho-c-Jun (S63) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-4, 1/50) for 30 minutes 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.
ET1608-4_8.jpg Fig8: 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) 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.
ET1608-4_9.jpg Fig9: 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) 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.