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Western blot analysis of extracts from NIH/3T3 and SK-N-MC cells, untreated or UV-treated, using c-Jun (60A8) Rabbit mAb.

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After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO* is added and emits light during enzyme catalyzed decomposition.

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After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

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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).

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Western blot analysis of extracts from untreated or anisomycin-treated C6 cells, or untreated or UV-treated 293 cells, using Phospho-c-Jun (Ser63) (54B3) Rabbit mAb (upper) or c-Jun (60A8) Rabbit mAb #9165 (lower).

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Immunohistochemical analysis of paraffin-embedded human astrocytoma, using c-Jun (60A8) Rabbit mAb.

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Immunohistochemical analysis of paraffin-embedded human breast carcinoma, control (left) or lambda phosphatase-treated (right), using Phospho-c-Jun (Ser73) (D47G9) XP® Rabbit mAb.

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Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Phospho-c-Jun (Ser63) (54B3) Rabbit mAb in the presence of control peptide (left) or Phospho-c-Jun (Ser63) II Blocking Peptide (#1020) (right).

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Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using c-Jun (60A8) Rabbit mAb.

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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).

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Immunohistochemical analysis of paraffin-embedded human breast carcinoma untreated (left) or lambda phosphatase-treated (right), using Phospho-c-Jun (Ser63) (54B3) Rabbit mAb.

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Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma, using c-Jun (60A8) Rabbit mAb.

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Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-c-Jun (Ser73) (D47G9) XP® Rabbit mAb.

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Immunohistochemical analysis of paraffin-embedded human lung carcinoma, showing nuclear localization, using Phospho-c-Jun (Ser63) (54B3) Rabbit mAb.

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Immunohistochemical analysis of frozen H1650 xenograft, showing nuclear localization using c-Jun (60A8) Rabbit mAb.

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Flow cytometric analysis of HeLa cells, untreated (blue) or UV treated (green), using Phospho-c-Jun (Ser73) (D47G9) XP® Rabbit mAb.

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Immunohistochemical analysis of paraffin-embedded NIH/3T3 cell pellets, control (left) or anisomycin-treated (right), using Phospho-c-Jun (Ser63) (54B3) Rabbit mAb.

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Confocal immunofluorescent analysis of HeLa cells, using c-Jun (60A8) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).

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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).

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Immunohistochemical analysis of paraffin-embedded human skin (normal adjacent to hemangioma), using Phospho-c-Jun (Ser63) (54B3) Rabbit mAb.

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Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 PC-12 cells starved overnight and treated with β-NGF #5221 (50ng/ml) for 2h and either 10 μl of c-Jun (60A8) Rabbit mAb or 2 μl of 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.

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Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 PC-12 cells starved overnight and treated with Human β-Nerve Growth Factor (hβ-NGF) #5221 (50 ng/ml) for 2h and either 10 μl of Phospho-c-Jun (Ser73) (D47G9) XP® Rabbit mAb or 2 μl of 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.

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Image
Product Includes Quantity Applications Reactivity MW(kDa) Isotype
c-Jun (60A8) Rabbit mAb 9165 x 100 µl
H M R Mk 43, 48 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 x 100 µl
All Goat 
Anti-biotin, HRP-linked Antibody 7075 x 100 µl
Goat 
20X LumiGLO® Reagent and 20X Peroxide 7003 x 5 ml each
 
Biotinylated Protein Ladder Detection Pack 7727 x 100 µl
 
c-Jun Control Cell Extracts 9263 x 150 µl
 
Phospho-c-Jun (Ser73) (D47G9) XP® Rabbit mAb 3270 x 100 µl
H M R Mk Pg 48 Rabbit IgG
Phospho-c-Jun (Ser63) (54B3) Rabbit mAb 2361 x 100 µl
H M R 48 Rabbit IgG

Product Description

The PhosphoPlus (R) c-Jun (Ser63) and c-Jun (Ser73) Antibody Kit provides reagents and controls for rapid analysis of c-Jun phosphorylation status.


Specificity / Sensitivity

Phospho-c-Jun (Ser63) (54B3) Rabbit mAb detects endogenous levels of c-Jun only when phosphorylated at Ser63. Phospho-c-Jun (Ser73) Antibody detects endogenous levels of c-Jun only when phosphorylated at Ser73. This antibody also recognizes phosphorylation of JunD at Ser100. c-Jun (60A8) Rabbit mAb detects endogenous levels of total c-Jun protein.


Source / Purification

Phospho-specific polyclonal antibodies are produced by immunizing rabbits with a synthetic phosphopeptide corresponding to residues surrounding Ser63 or Ser73 of human c-Jun, and purified by protein A and peptide affinity chromatography. Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the amino-terminal of human c-Jun.

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). Knock-out 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.


Entrez-Gene Id 3725
Swiss-Prot Acc. P05412


For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
PhosphoPlus® is a trademark of Cell Signaling Technology, Inc.
DRAQ5® is a registered trademark of Biostatus Limited.
LumiGLO® is a registered trademark of Kirkegaard & Perry Laboratories.
U.S. Patent No. 5,675,063.