Revision 1
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
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UniProt ID:

#P05412

Entrez-Gene Id:

3725

Product Includes Quantity Reactivity MW(kDa) Isotype
Phospho-c-Jun (Ser73) (D47G9) XP® Rabbit mAb 3270 100 µl H M R Mk Pg 48 Rabbit IgG
c-Jun (60A8) Rabbit mAb 9165 100 µl H M R Mk 43, 48 Rabbit IgG

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

PhosphoPlus® Duets from Cell Signaling Technology (CST) provide a means to assess protein activation status. Each Duet contains an activation-state and total protein antibody to your target of interest. These antibodies have been selected from CST's product offering based upon superior performance in specified applications.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Background

c-Jun is a member of the Jun family containing c-Jun, JunB, and JunD, and is a component of the transcription factor activator protein-1 (AP-1). AP-1 is composed of dimers of Fos, Jun, and ATF family members and binds to and activates transcription at TRE/AP-1 elements (reviewed in 1). Extracellular signals, including growth factors, chemokines, and stress, activate AP-1-dependent transcription. The transcriptional activity of c-Jun is regulated by phosphorylation at Ser63 and Ser73 through SAPK/JNK (reviewed in 2). Knockout studies in mice have shown that c-Jun is essential for embryogenesis (3), and subsequent studies have demonstrated roles for c-Jun in various tissues and developmental processes, including axon regeneration (4), liver regeneration (5), and T cell development (6). AP-1 regulated genes exert diverse biological functions, including cell proliferation, differentiation, and apoptosis, as well as transformation, invasion and metastasis, depending on cell type and context (7-9). Other target genes regulate survival, as well as hypoxia and angiogenesis (8,10). Research studies have implicated c-Jun as a promising therapeutic target for cancer, vascular remodeling, acute inflammation, and rheumatoid arthritis (11,12).

  1. Jochum, W. et al. (2001) Oncogene 20, 2401-12.
  2. Davis, R.J. (2000) Cell 103, 239-52.
  3. Hilberg, F. et al. (1993) Nature 365, 179-81.
  4. Raivich, G. et al. (2004) Neuron 43, 57-67.
  5. Behrens, A. et al. (2002) EMBO J 21, 1782-90.
  6. Riera-Sans, L. and Behrens, A. (2007) J Immunol 178, 5690-700.
  7. Leppä, S. and Bohmann, D. (1999) Oncogene 18, 6158-62.
  8. Shaulian, E. and Karin, M. (2002) Nat Cell Biol 4, E131-6.
  9. Weiss, C. and Bohmann, D. (2004) Cell Cycle 3, 111-3.
  10. Karamouzis, M.V. et al. (2007) Mol Cancer Res 5, 109-20.
  11. Kim, S. and Iwao, H. (2003) J Pharmacol Sci 91, 177-81.
  12. Dass, C.R. and Choong, P.F. (2008) Pharmazie 63, 411-4.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    PhosphoPlus is a registered trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    使用に関する制限

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    Revision 1
    #8222

    PhosphoPlus® c-Jun (Ser73) Antibody Duet

    PhosphoPlus® c-Jun (Ser73) Antibody Duet: Image 1 Expand Image

    Simple Western™ analysis of lysates (0.1 mg/mL) from HeLa cells treated with UV (100 mJ/cm2; 2H recovery) using c-Jun (60A8) Rabbit mAb #9165. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ ​​​​​​​ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.

    PhosphoPlus® c-Jun (Ser73) Antibody Duet: Image 2 Expand Image
    Western blot analysis of extracts from NIH/3T3 or C6 cells, untreated or UV-treated, using Phospho-c-Jun (Ser73) (D47G9) XP® Rabbit mAb (upper) or c-Jun (60A8) Rabbit mAb #9165 (lower).
    PhosphoPlus® c-Jun (Ser73) Antibody Duet: Image 3 Expand Image
    CUT&RUN was performed with PC-12 cells starved overnight and treated with Human β-Nerve Growth Factor (hβ-NGF) #5221 (50 ng/ml) for 2h and Phospho-c-Jun (Ser73) (D47G9) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Library was prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across Lmna, a known target gene of Phospho-c-Jun (see additional figure containing CUT&RUN-qPCR data).
    PhosphoPlus® c-Jun (Ser73) Antibody Duet: Image 4 Expand Image
    Western blot analysis of extracts from control HeLa cells (lane 1) or c-Jun knockout HeLa cells (lane 2) using c-Jun (60A8) Rabbit mAb #9165. The absence of signal in the c-Jun knockout HeLa cells confirms specificity of the antibody for c-Jun.
    PhosphoPlus® c-Jun (Ser73) Antibody Duet: Image 5 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma, control (left) or lambda phosphatase-treated (right), using Phospho-c-Jun (Ser73) (D47G9) XP® Rabbit mAb.
    PhosphoPlus® c-Jun (Ser73) Antibody Duet: Image 6 Expand Image
    CUT&RUN was performed with PC-12 cells starved overnight and treated with Human β-Nerve Growth Factor (hβ-NGF) #5221 (50 ng/ml) for 2h and Phospho-c-Jun (Ser73) (D47G9) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Library was prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 2 (upper), including Lmna (lower), a known target gene of Phospho-c-Jun (see additional figure containing CUT&RUN-qPCR data).
    PhosphoPlus® c-Jun (Ser73) Antibody Duet: Image 7 Expand Image
    Western blot analysis of extracts from NIH/3T3 and SK-N-MC cells, untreated or UV-treated, using c-Jun (60A8) Rabbit mAb.
    PhosphoPlus® c-Jun (Ser73) Antibody Duet: Image 8 Expand Image
    Immunohistochemical analysis of parafin-embedded human colon carcinoma using Phospho-c-Jun (Ser73) (D47G9) XP® Rabbit mAb in the presence of control peptide (left) or Phospho-c-Jun (Ser73) Blocking Peptide (right).
    PhosphoPlus® c-Jun (Ser73) Antibody Duet: Image 9 Expand Image
    CUT&RUN was performed with PC-12 cells starved overnight and treated with Human β-Nerve Growth Factor (hβ-NGF) #5221 (50 ng/ml) for 2h and either Phospho-c-Jun (Ser73) (D47G9) XP® Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using rat Lmna promoter primer, rat Cic intron 1 primer and rat Phf17 promoter primer. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
    PhosphoPlus® c-Jun (Ser73) Antibody Duet: Image 10 Expand Image
    Immunohistochemical analysis of paraffin-embedded human astrocytoma, using c-Jun (60A8) Rabbit mAb.
    PhosphoPlus® c-Jun (Ser73) Antibody Duet: Image 11 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-c-Jun (Ser73) (D47G9) XP® Rabbit mAb.
    PhosphoPlus® c-Jun (Ser73) Antibody Duet: Image 12 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using c-Jun (60A8) Rabbit mAb.
    PhosphoPlus® c-Jun (Ser73) Antibody Duet: Image 13 Expand Image
    Confocal immunofluorescent analysis of mouse small intestine, untreated (left) or treated with λ-phosphatase (right), using Phospho-c-Jun (Ser73) (D47G9) XP® Rabbit mAb #3270 (green). Actin filaments have been labeled with DY-554 phalloidin (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    PhosphoPlus® c-Jun (Ser73) Antibody Duet: Image 14 Expand Image
    Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma, using c-Jun (60A8) Rabbit mAb.
    PhosphoPlus® c-Jun (Ser73) Antibody Duet: Image 15 Expand Image
    Confocal immunofluorescent analysis of HeLa cells, untreated (left) or anisomycin-treated (right), using Phospho-c-Jun (Ser73) (D47G9) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).
    PhosphoPlus® c-Jun (Ser73) Antibody Duet: Image 16 Expand Image
    Flow cytometric analysis of HeLa cells, untreated (blue) or UV treated (green), using Phospho-c-Jun (Ser73) (D47G9) XP® Rabbit mAb.
    PhosphoPlus® c-Jun (Ser73) Antibody Duet: Image 17 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from PC-12 cells starved overnight and treated with Human β-Nerve Growth Factor (hβ-NGF) #5221 (50 ng/ml) for 2h and either Phospho-c-Jun (Ser73) (D47G9) XP® Rabbit mAb or c-Jun (60A8) Rabbit mAb #9165, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across Dclk1, a known target gene of both Phospho-c-Jun and c-Jun (see additional figures containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product datasheet.
    PhosphoPlus® c-Jun (Ser73) Antibody Duet: Image 18 Expand Image
    Confocal immunofluorescent analysis of HeLa cells, using c-Jun (60A8) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).
    PhosphoPlus® c-Jun (Ser73) Antibody Duet: Image 19 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from PC-12 cells starved overnight and treated with Human β-Nerve Growth Factor (hβ-NGF) #5221 (50 ng/ml) for 2h and either Phospho-c-Jun (Ser73) (D47G9) XP® Rabbit mAb or c-Jun (60A8) Rabbit mAb #9165, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 2 (upper), including Dclk1 (lower), a known target gene of both Phospho-c-Jun and c-Jun (see additional figures containing ChIP-qPCR data).
    PhosphoPlus® c-Jun (Ser73) Antibody Duet: Image 20 Expand Image
    Flow cytometric analysis of Jurkat cells using c-Jun (60A8) Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
    PhosphoPlus® c-Jun (Ser73) Antibody Duet: Image 21 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from cells starved overnight and treated with Human β-Nerve Growth Factor (hβ-NGF) #5221 (50 ng/ml) for 2h and either Phospho-c-Jun (Ser73) (D47G9) XP® Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR SimpleChIP® using Rat CCRN4L Promoter Primers #7983, rat DCLK1 promoter primers, and SimpleChIP® Rat GAPDH Promoter Primers #7964. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
    PhosphoPlus® c-Jun (Ser73) Antibody Duet: Image 22 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from PC-12 cells starved overnight and treated with β-NGF #5221 (50ng/ml) for 2h and c-Jun (60A8) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across Dclk1, a known target gene of c-Jun (see additional figure containing ChIP-qPCR data).
    PhosphoPlus® c-Jun (Ser73) Antibody Duet: Image 23 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from PC-12 cells starved overnight and treated with β-NGF #5221 (50ng/ml) for 2h and c-Jun (60A8) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 2 (upper), including Dclk1 (lower), a known target gene of c-Jun (see additional figure containing ChIP-qPCR data).
    PhosphoPlus® c-Jun (Ser73) Antibody Duet: Image 24 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from PC-12 cells starved overnight and treated with β-NGF #5221 (50ng/ml) for 2h and either of c-Jun (60A8) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Rat CCRN4L Promoter Primers #7983, rat DCLK1 promoter primers, and SimpleChIP® Rat GAPDH Promoter Primers #7964. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.