Cell Signaling Technology

Product Pathways - MAPK Signaling

PhosphoPlus® c-Jun (Ser63) and c-Jun (Ser73) Antibody Kit #9260

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

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IHC-F=Immunohistochemistry (Frozen)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

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.

Western Blotting

Western Blotting

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 #2361 (upper) or c-Jun (60A8) Rabbit mAb #9165 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from UV-treated NIH/3T3 cells using Phospho-c-Jun (Ser73) Antibody #9164.

Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3 and SK-N-MC cells, untreated or UV-treated, using c-Jun (60A8) Rabbit mAb #9165.


IHC-F (frozen)

IHC-F (frozen)

Immunohistochemical analysis of frozen H1650 xenograft, showing nuclear localization using c-Jun (60A8) Rabbit mAb #9165.

IF-IC

IF-IC

Confocal immunofluorescent images of C6 cells, untreated (left) or anisomycin treated (right), labeled with Phospho-c-Jun (Ser73) Antibody #9164 (green) and b-Tubulin Antibody #2146 (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, using c-Jun (60A8) Rabbit mAb #9165 (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).


Description

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

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.

Background

c-Jun is a member of the Jun Family containing c-Jun, JunB and JunD, and is a component of the transcription factor AP-1 (activator protein-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). c-Jun has emerged as a promising therapeutic target for cancer, vascular remodeling, acute inflammation, as well as 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.

Application References

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This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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