Cell Signaling Technology

Product Pathways - MAPK Signaling

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

Kit Includes Quantity Applications Reactivity MW (kDa) Source
Phospho-c-Jun (Ser63) II Antibody # 9261 100 microliters W F H M R Pg 48 Rabbit
Phospho-c-Jun (Ser73) Antibody # 9164 100 microliters W IHC-P IF-IC H M R 48 Rabbit
c-Jun (60A8) Rabbit mAb # 9165 100 microliters W IP IHC-P IHC-F IF-IC H M R Mk 43, 48 Rabbit
Anti-rabbit IgG, HRP-linked Antibody # 7074 50 microliters Goat
Anti-biotin, HRP-linked Antibody # 7075 100 microliters Goat
20X LumiGLO® Reagent and 20X Peroxide # 7003 5 milliliters
Biotinylated Protein Ladder Detection Pack # 7727 100 microliters
c-Jun Control Cell Extracts # 9263 60 microliters

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  Pg=Pig

Specificity / Sensitivity

Phospho-c-Jun (Ser 63) II Antibody detects endogenous levels of c-Jun only when phosphorylated at Ser 63. This antibody does not recognize the phosphorylated forms of JunD or JunB. Phospho-c-Jun (Ser 73) Antibody detects endogenous levels of c-Jun only when phosphorylated at Ser 73. This antibody also recognizes phosphorylation of JunD at Ser 100. c-Jun (60A8) Rabbit mAb detects endogenous levels of total c-Jun protein.

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.

Western Blotting

Western Blotting

Western Analysis of extracts from anisomycin or UV-treated NIH/3T3 cells using Phospho-c-Jun (Ser63) II Antibody #9261.


IHC-F (frozen)

IHC-F (frozen)

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

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (blue) or Anisomycin-treated (green), using Phospho-c-Jun (Ser63) II Antibody #9261 compared to a nonspecific negative control antibody (red).

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).

Source / Purification

Phospho-specific polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Ser63 or Ser73 of human c-Jun, and purified by protein A and peptide affinity chromatography. Rabbit monoclonal antibody is produced by immunizing rabbits with a synthetic peptide (KLH-coupled) derived from the amino-terminal of human c-Jun.

Background

c-Jun is a component of the transcription factor AP-1, which binds and activates transcription at TRE/AP-1 elements. The transcriptional activity of c-Jun is regulated by phosphorylation at Ser63 and Ser73 (1,2). Extracellular signals including growth factors, transforming oncoproteins and UV irradiation stimulate phosphorylation of c-Jun at Ser63/73 and activate c-Jun-dependent transcription. Mutation of Ser63/73 renders c-Jun nonresponsive to mitogenic and stress-induced signaling pathways. The MAP kinase homologue SAPK/JNK binds to the amino-terminal region of c-Jun and phosphorylates c-Jun at Ser63/73. In addition, the activity of SAPK/JNK is stimulated by the same signals that activate c-Jun (3,4).

  1. Binetruy, B. et al. (1991) Nature 351, 122-127.
  2. Smeal, T. et al. (1991) Nature 354, 494-496.
  3. Derijard, B. et al. (1994) Cell 76, 1025-1037.
  4. Kyriakis, J.M. et al. (1994) Nature 369, 156-160.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

Product Pathways

Drug Discovery Tools

Featured Technologies

Protein Classes