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Phospho-c-Jun (S73)+JunD (S100) Recombinant Rabbit Monoclonal Antibody [JE65-46]

Usd: 385

Specification

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Catalog# HA722475

Phospho-c-Jun (S73)+JunD (S100) Recombinant Rabbit Monoclonal Antibody [JE65-46]

Application

  • WB

  • IF-Cell

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

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

Overview

Product Name

Phospho-c-Jun (S73)+JunD (S100) Recombinant Rabbit Monoclonal Antibody [JE65-46]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic phosphopeptide corresponding to residues around Ser73 of human c-Jun.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, FC

Molecular Weight

Predicted band size: 40/45 kDa

Positive Control

HeLa cell lysate, HeLa treated with 25μg/mL anisomycin for 30 minutes cell lysate, NIH/3T3 cell lysate, NIH/3T3 treated with 25μg/mL anisomycin for 30 minutes cell lysate, RAW264.7 cell lysate, PC-12 cell lysate.

Conjugation

unconjugated

Clone Number

JE65-46

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

  • 1:1,000

  • IF-Cell

  • 1:100

  • FC

  • 1:1,000

Target

Function

Transcription factor that recognizes and binds to the AP-1 consensus motif 5'-TGA[GC]TCA-3'. Heterodimerizes with proteins of the FOS family to form an AP-1 transcription complex, thereby enhancing its DNA binding activity to the AP-1 consensus sequence 5'-TGA[GC]TCA-3' and enhancing its transcriptional activity. Together with FOSB, plays a role in activation-induced cell death of T cells by binding to the AP-1 promoter site of FASLG/CD95L, and inducing its transcription in response to activation of the TCR/CD3 signaling pathway. Promotes activity of NR5A1 when phosphorylated by HIPK3 leading to increased steroidogenic gene expression upon cAMP signaling pathway stimulation. Involved in activated KRAS-mediated transcriptional activation of USP28 in colorectal cancer (CRC) cells. (Microbial infection) Upon Epstein-Barr virus (EBV) infection, binds to viral BZLF1 Z promoter and activates viral BZLF1 expression.

Background References

1. Qing J., Zhang Y., Derynck R. Structural and functional characterization of the transforming growth factor-beta -induced Smad3/c-Jun transcriptional cooperativity. J. Biol. Chem. 275:38802-38812 (2000)

2. Serra R.W., Fang M., Park S.M., Hutchinson L., Green M.R. A KRAS-directed transcriptional silencing pathway that mediates the CpG island methylator phenotype. Elife 3:E02313-E02313 (2014)

Subcellular Location

Nucleus.

UNIPROT

SwissProt: P05412 Human

SwissProt: P17535 Human

SwissProt: P05627 Mouse

SwissProt: P15066 Mouse

SwissProt: P17325 Rat

SwissProt: P52909 Rat

Synonyms

Activator protein 1 antibody

AP 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

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-c-Jun (S73)+JunD (S100) on different lysates with Rabbit anti-Phospho-c-Jun (S73)+JunD (S100) antibody (<a href="/products/HA722475" style="font-weight: bold;text-decoration: underline;">HA722475</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HeLa treated with 25μg/mL anisomycin for 30 minutes cell lysate<br />Lane 3: HeLa treated with 25μg/mL anisomycin for 30 minutes cell lysate, then the membrane treated with λpp for 1 hour<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 40/45 kDa<br />Observed band size: 40/45 kDa<br /><br />Exposure time: 30 seconds; ECL: K1801;<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/HA722475" style="font-weight: bold;text-decoration: underline;">HA722475</a>) at 1/1,000 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 (S73)+JunD (S100) on different lysates with Rabbit anti-Phospho-c-Jun (S73)+JunD (S100) antibody (HA722475) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HeLa treated with 25μg/mL anisomycin for 30 minutes cell lysate
    Lane 3: HeLa treated with 25μg/mL anisomycin for 30 minutes cell lysate, then the membrane treated with λpp for 1 hour

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 40/45 kDa
    Observed band size: 40/45 kDa

    Exposure time: 30 seconds; ECL: K1801;
    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 (HA722475) at 1/1,000 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.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-c-Jun (S73)+JunD (S100) on different lysates with Rabbit anti-Phospho-c-Jun (S73)+JunD (S100) antibody (<a href="/products/HA722475" style="font-weight: bold;text-decoration: underline;">HA722475</a>) at 1/1,000 dilution.<br /><br />Lane 1: NIH/3T3 cell lysate<br />Lane 2: NIH/3T3 treated with 25μg/mL anisomycin for 30 minutes cell lysate<br />Lane 3: RAW264.7 cell lysate<br />Lane 4: PC-12 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 40/45 kDa<br />Observed band size: 40/45 kDa<br /><br />Exposure time: 3 minutes; ECL: K1801;<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/HA722475" style="font-weight: bold;text-decoration: underline;">HA722475</a>) at 1/1,000 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 (S73)+JunD (S100) on different lysates with Rabbit anti-Phospho-c-Jun (S73)+JunD (S100) antibody (HA722475) at 1/1,000 dilution.

    Lane 1: NIH/3T3 cell lysate
    Lane 2: NIH/3T3 treated with 25μg/mL anisomycin for 30 minutes cell lysate
    Lane 3: RAW264.7 cell lysate
    Lane 4: PC-12 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 40/45 kDa
    Observed band size: 40/45 kDa

    Exposure time: 3 minutes; ECL: K1801;
    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 (HA722475) at 1/1,000 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.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Immunocytochemistry analysis of HeLa cells untreated / treated with 250ng/mL anisomycin for 30 minutes labeling Phospho-c-Jun (S73)+JunD (S100) with Rabbit anti-Phospho-c-Jun (S73)+JunD (S100) antibody (<a href="/products/HA722475" style="font-weight: bold;text-decoration: underline;">HA722475</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-c-Jun (S73)+JunD (S100) antibody (<a href="/products/HA722475" style="font-weight: bold;text-decoration: underline;">HA722475</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    ☑ Cell treatment (CT)

    Immunocytochemistry analysis of HeLa cells untreated / treated with 250ng/mL anisomycin for 30 minutes labeling Phospho-c-Jun (S73)+JunD (S100) with Rabbit anti-Phospho-c-Jun (S73)+JunD (S100) antibody (HA722475) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-c-Jun (S73)+JunD (S100) antibody (HA722475) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Flow cytometric analysis of HeLa cells untreated (left) / treated with 250ng/mL anisomycin for 30 minutes (right) labeling Phospho-c-Jun (S73)+JunD (S100).<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA722475" style="font-weight: bold;text-decoration: underline;">HA722475</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    ☑ Cell treatment (CT)

    Flow cytometric analysis of HeLa cells untreated (left) / treated with 250ng/mL anisomycin for 30 minutes (right) labeling Phospho-c-Jun (S73)+JunD (S100).

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA722475, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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

Citation

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metafield image

Phospho-c-Jun (S73)+JunD (S100) Recombinant Rabbit Monoclonal Antibody [PSH07-12] - BSA and Azide free

Application: WB,IF-Cell

Reactivity: Human,Mouse,Rat

Conjugate: unconjugated

metafield image

Phospho-c-Jun (S73)+JunD (S100) Recombinant Rabbit Monoclonal Antibody [PSH07-12]

Application: WB,IF-Cell

Reactivity: Human,Mouse,Rat

Conjugate: unconjugated