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 blot analysis of extracts from UV-treated NIH/3T3 cells using Phospho-c-Jun (Ser73) Antibody #9164.
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 Analysis of extracts from anisomycin or UV-treated NIH/3T3 cells using Phospho-c-Jun (Ser63) II Antibody #9261.
IHC-F (frozen)
Immunohistochemical analysis of frozen H1650 xenograft, showing nuclear localization using c-Jun (60A8) Rabbit mAb #9165.
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
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
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).
- Binetruy, B. et al. (1991) Nature 351, 122-127.
- Smeal, T. et al. (1991) Nature 354, 494-496.
- Derijard, B. et al. (1994) Cell 76, 1025-1037.
- Kyriakis, J.M. et al. (1994) Nature 369, 156-160.
Application References
- Li, B. S. et al. (2002) Cyclin-dependent kinase 5 prevents neuronal apoptosis by negative regulation of c-Jun N-terminal kinase 3. EMBO J. 21, 324-333. This article references the use of PhosphoPlus® c-Jun (Ser63) II and c-Jun (Ser73) Antibody Kit in the following applications: IC-ABC Western Blotting
- Garcia, M. et al. (2002) The mitochondrial toxin 3-nitropropionic acid induces striatal neurodegeneration via a c-Jun N-terminal kinase/c-Jun module. J. Neurosci. 22, 2174-2184. This article references the use of PhosphoPlus® c-Jun (Ser63) II and c-Jun (Ser73) Antibody Kit in the following applications: IHC-FL (floating/frozen) Western Blotting
- Watanabe, H. et al. (2001) Transcriptional cross-talk between Smad, ERK1/2, and p38 mitogen-activated protein kinase pathways regulates transforming growth factor-beta-induced aggrecan gene expression in chondrogenic ATDC5 cells. J. Biol. Chem. 276, 14466-14473. This article references the use of PhosphoPlus® c-Jun (Ser63) II and c-Jun (Ser73) Antibody Kit in the following applications: Western Blotting
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Companion Products
- 9151 Phospho-SEK1/MKK4 (Thr261) Antibody
- 9810 SAPK/JNK Kinase Assay Kit (Nonradioactive)
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 9251 Phospho-SAPK/JNK (Thr183/Tyr185) Antibody
- 9255 Phospho-SAPK/JNK (Thr183/Tyr185) (G9) Mouse mAb
- 9258 SAPK/JNK (56G8) Rabbit mAb
- 9252 SAPK/JNK Antibody
- 9221 Phospho-ATF-2 (Thr71) Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7727 Biotinylated Protein Ladder Detection Pack
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7003 20X LumiGLO® Reagent and 20X Peroxide
