Phospho-c-Jun (T91) Recombinant Rabbit Monoclonal Antibody [JJ080-9]
cat.: ET1701-32
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, IP
Clonality: Monoclonal
Clone number: JJ080-9
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 Thr91 of Human c-Jun aa 60-109 / 331.
Positive control: 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, NIH/3T3 treated with UV for 15 minutes then recover 30 minutes whole cell lysate, HepG2, MCF-7, human breast carcinoma tissue, human skin tissue, human breast tissue.
Subcellular location: Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  IP
1:500
1:50-1:200
1:50-1:200
1:50-1:200
Use at an assay dependent concentration.
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
ET1701-32_1.jpg Fig1: Western blot analysis of Phospho-c-Jun (T91) on different lysates with Rabbit anti-Phospho-c-Jun (T91) antibody (ET1701-32) at 1/500 dilution.

Lane 1: A549 whole cell lysate (20 µg/Lane)
Lane 2: A549 treated with 250ng/mL anisomycin for 30 minutes whole cell lysate (20 µg/Lane)
Lane 3: NIH/3T3 whole cell lysate (20 µg/Lane)
Lane 4: NIH/3T3 treated with 250ng/mL anisomycin for 30 minutes whole cell lysate (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 (ET1701-32) 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.
ET1701-32_2.jpg Fig2: Western blot analysis of Phospho-c-Jun (T91) on different lysates with Rabbit anti-Phospho-c-Jun (T91) antibody (ET1701-32) at 1/500 dilution.

Lane 1: NIH/3T3 whole cell lysate
Lane 2: NIH/3T3 treated with UV for 15 minutes then recover 30 minutes whole cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 36 kDa
Observed band size: 40 kDa

Exposure time: 1 minute 30 seconds;

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 (ET1701-32) 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 1:200,000 dilution was used for 1 hour at room temperature.
ET1701-32_3.jpg Fig3: ICC staining of Phospho-c-Jun (T91) in HepG2 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 (ET1701-32, 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).
ET1701-32_4.jpg Fig4: ICC staining of Phospho-c-Jun (T91) 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 (ET1701-32, 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).
ET1701-32_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-Phospho-c-Jun (T91) 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 (ET1701-32, 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.
ET1701-32_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human skin tissue using anti-Phospho-c-Jun (T91) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (ET1701-32, 1/200) 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.
ET1701-32_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human breast tissue using anti-Phospho-c-Jun (T91) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (ET1701-32, 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.