PRODUCT CODE: ET1608-4

Phospho-c-Jun (S63) Recombinant Rabbit Monoclonal Antibody [SY0297] (ET1608-4)

  • Recombinant

Applications

  • WB

  • ICC

  • IHC-P

REACTIVITY

  • Human

  • Mouse

  • Rat

Western blot analysis of Phospho-c-Jun(S63) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1608-4, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: 293T cell lysate<br />
Lane 2: NIH/3T3 cell lysate
  • Western blot analysis of Phospho-c-Jun(S63) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1608-4, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: 293T cell lysate<br />
Lane 2: NIH/3T3 cell lysate
  • 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).
  • 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).
  • 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).
  • 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.
  • 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.
Western blot analysis of Phospho-c-Jun(S63) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1608-4, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: 293T cell lysate
Lane 2: NIH/3T3 cell lysate

Applications

  • WB

  • ICC

  • IHC-P

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Phospho-c-Jun (S63) Recombinant Rabbit Monoclonal Antibody [SY0297] (ET1608-4)

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding ser63 of human c-jun.

Host

Rabbit

Modification

Phospho

Modification Site

S63

Positive Control

293T cell lysate, NIH/3T3 cell lysate, PC-3M, MCF-7, A549, human breast carcinoma tissue, human tonsil tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SY0297

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

40 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000-1:2,000

  • ICC

  • 1:50-1:200

  • IHC-P

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

Phospho-c-Jun (S63)

SYNONYMS

Activator protein 1 antibody; AP 1 antibody; AP1 antibody; cJun antibody; Enhancer Binding Protein AP1 antibody; Jun Activation Domain Binding Protein antibody; JUN antibody; Jun oncogene antibody; JUN protein antibody; Jun proto oncogene antibody; JUN_HUMAN antibody; JUNC antibody; Oncogene JUN antibody; p39 antibody; Proto oncogene c jun antibody; Proto oncogene cJun antibody; Proto-oncogene c-jun antibody; Transcription Factor AP 1 antibody; Transcription factor AP-1 antibody; Transcription Factor AP1 antibody; V jun avian sarcoma virus 17 oncogene homolog antibody; V jun sarcoma virus 17 oncogene homolog (avian) antibody; V jun sarcoma virus 17 oncogene homolog antibody; V-jun avian sarcoma virus 17 oncogene homolog antibody; vJun Avian Sarcoma Virus 17 Oncogene Homolog antibody

SEQUENCE SIMILARITIES

Belongs to the bZIP family. Jun subfamily.

TISSUE SPECIFICITY

Expressed in the developing and adult prostate and prostate cancer cells.

POST-TRANSLATIONAL MODIFICATION

Ubiquitinated by the SCF(FBXW7), leading to its degradation. Ubiquitination takes place following phosphorylation, that promotes interaction with FBXW7.; Phosphorylated by CaMK4 and PRKDC; phosphorylation enhances the transcriptional activity. Phosphorylated by HIPK3. Phosphorylated by DYRK2 at Ser-243; this primes the protein for subsequent phosphorylation by GSK3B at Thr-239. Phosphorylated at Thr-239, Ser-243 and Ser-249 by GSK3B; phosphorylation reduces its ability to bind DNA. Phosphorylated by PAK2 at Thr-2, Thr-8, Thr-89, Thr-93 and Thr-286 thereby promoting JUN-mediated cell proliferation and transformation. Phosphorylated by PLK3 following hypoxia or UV irradiation, leading to increase DNA-binding activity.; Acetylated at Lys-271 by EP300.

SUBCELLULAR LOCATION

Nucleus.

FUNCTION

Genes belonging to the Jun and Fos oncogene families encode nuclear proteins that are associated with a number of transcriptional complexes. The c-Jun protein is a major component of the transcription factor AP-1, originally shown to mediate phorbol ester tumor promoter (TPA)-induced expression of responsive genes through the TPA-response element (TRE). The Jun proteins form homo- and heterodimers which bind the TRE, while Fos proteins are active only as heterodimers with any of the Jun proteins. Fos/Jun heterodimers have a much higher affinity for the TRE than Jun homodimers. Ha-Ras augments c-Jun activity and stimulates phosphorylation of its activation domain. An inhibitor of Fos/Jun function, termed IP-1, associates with Fos and Jun and is inactivated upon phosphorylation induced by the cAMP-dependent protein kinase A (PKA).