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

Usd: 385

Specification

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Catalog# ET1608-4

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

Application

  • WB

  • IF-Cell

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

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

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding Ser63 of Human c-Jun aa 31-80 / 331.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P

Molecular Weight

Predicted band size: 36 kDa

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.

Conjugation

unconjugated

Clone Number

SY0297

RRID

AB_3069804

Product Features

Form

Liquid

Concentration

Storage Instructions

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.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:500-1:2,000

  • IF-Cell

  • 1:50-1:200

  • IHC-P

  • 1:500

Target

Function

c-Jun is a protein that in humans is encoded by the JUN gene. c-Jun, in combination with c-Fos, forms the AP-1 early response transcription factor. It was first identified as the Fos-binding protein p39 and only later rediscovered as the product of the JUN gene. c-jun was the first oncogenic transcription factor discovered. The proto-oncogene c-Jun is the cellular homolog of the viral oncoprotein v-jun (P05411). The viral homolog v-jun was discovered in avian sarcoma virus 17 and was named for ju-nana, the Japanese word for 17. The human JUN encodes a protein that is highly similar to the viral protein, which interacts directly with specific target DNA sequences to regulate gene expression. This gene is intronless and is mapped to 1p32-p31, a chromosomal region involved in both translocations and deletions in human malignancies.

Background References

1. Zhang QS et al. Downregulation of SENP1 inhibits cell proliferation, migration and promotes apoptosis in human glioma cells. Oncol Lett 12:217-221 (2016).

2. Li C et al. Inhibitory effects of kaempferol on the invasion of human breast carcinoma cells by downregulating the expression and activity of matrix metalloproteinase-9. Biochem Cell Biol 93:16-27 (2015).

Sequence Similarity

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.

UNIPROT

SwissProt: P05412 Human

SwissProt: P05627 Mouse

SwissProt: P17325 Rat

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

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-c-Jun (S63) on different lysates with Rabbit anti-Phospho-c-Jun (S63) antibody (<a href="/products/ET1608-4" style="font-weight: bold;text-decoration: underline;">ET1608-4</a>) at 1/2,000 dilution.<br /><br />Lane 1: C6 cell lysate<br />Lane 2: C6 treated with 25μg/mL Anisomycin for 30 minutes cell lysate<br />Lane 3: NIH/3T3 cell lysate<br />Lane 4: NIH/3T3 treated with 25μg/mL Anisomycin for 30 minutes cell lysate<br />Lane 5: NIH/3T3 treated with 25μg/mL Anisomycin for 30 minutes, then treated with λpp for 1 hour cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 36 kDa<br />Observed band size: 40 kDa<br /><br />Exposure time: 1 minute;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1608-4" style="font-weight: bold;text-decoration: underline;">ET1608-4</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    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.

  • 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 (<a href="/products/ET1608-4" style="font-weight: bold;text-decoration: underline;">ET1608-4</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;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 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).

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-c-Jun (S63) on different lysates with Rabbit anti-Phospho-c-Jun (S63) antibody (<a href="/products/ET1608-4" style="font-weight: bold;text-decoration: underline;">ET1608-4</a>) at 1/500 dilution.<br /><br />Lane 1: A549 whole cell lysate<br />Lane 2: A549 treated with 250ng/mL anisomycin for 30 minutes whole cell lysate<br />Lane 3: NIH/3T3 whole cell lysate<br />Lane 4: NIH/3T3 treated with 250ng/mL anisomycin for 30 minutes whole cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 36 kDa<br />Observed band size: 40 kDa<br /><br />Exposure time: 5 minutes;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1608-4" style="font-weight: bold;text-decoration: underline;">ET1608-4</a>) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:100,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    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.

  • 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 (<a href="/products/ET1608-4" style="font-weight: bold;text-decoration: underline;">ET1608-4</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;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 (<a href="/products/ET1608-4" style="font-weight: bold;text-decoration: underline;">ET1608-4</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;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 colon carcinoma tissue with Rabbit anti-Phospho-c-Jun (S63) antibody (<a href="/products/ET1608-4" style="font-weight: bold;text-decoration: underline;">ET1608-4</a>) at 1/500 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1608-4" style="font-weight: bold;text-decoration: underline;">ET1608-4</a>) 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.

    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.

  • Immunohistochemical analysis of paraffin-embedded human endometrial tissue with Rabbit anti-Phospho-c-Jun (S63) antibody (<a href="/products/ET1608-4" style="font-weight: bold;text-decoration: underline;">ET1608-4</a>) at 1/500 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1608-4" style="font-weight: bold;text-decoration: underline;">ET1608-4</a>) 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.

    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.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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