Product Pathways - PathScan ELISA

PathScan® Phospho-c-Jun (Ser63) Sandwich ELISA Kit II #7145

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Kit Includes	Volume	Solution Color
Phospho-c-Jun (Ser63) Rabbit mAb Coated Microwells
c-Jun Detection Antibody	11 milliliters
Anti-mouse IgG HRP-Linked Antibody	11 milliliters
TMB Substrate	11 milliliters	Colorless
STOP Solution	11 milliliters	Colorless
Sealing Tape	2 sheets
20X Wash Buffer	25 milliliters	Colorless
Sample Diluent	25 milliliters	Blue
Cell Lysis Buffer (10X) # 9803	15 milliliters	Yellowish

Note: 12 8-well modules –Each module is designed to break apart for 8 tests.
Note: Kit should be stored at 4°C with the exception of Cell Lysis Buffer (10X), which is stored at –20°C (packaged separately).

Species Cross-Reactivity

H M R

Reactivity Key:  H=Human  M=Mouse  R=Rat

Description

CST's PathScan® Phospho-c-Jun (Ser63) Sandwich ELISA Kit II is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-c-Jun (Ser63) protein. A phospho-c-Jun (Ser63)-specific rabbit mAb (#2378*) has been coated onto the microwells. After incubation with cell lysates, phospho-c-Jun (Ser63) protein is captured by the coated antibody. Following extensive washing, c-Jun Mouse mAb (#2379*) is added to detect the captured phospho-c-Jun protein. HRP-linked anti-mouse antibody (#7076*) is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of phospho-c-Jun (Ser63) protein.* Antibodies in this kit are custom formulations specific to the kit.

Specificity / Sensitivity

PathScan® Phospho-c-Jun (Ser63) Sandwich ELISA Kit II detects endogenous levels of Phospho-c-Jun (Ser63) protein. As shown in Figure 1, using this Sandwich ELISA Kit #7145, a significant induction of phospho-c-Jun (Ser63) in UV-treated 293 cells can be detected. However, the level of total c-Jun protein (phospho and non-phospho), detected by the Total c-Jun Sandwich ELISA Kit II #7150, remains unchanged. Both C6 and NIH/3T3 cells treated with UV light or anisomycin show similar results (data not shown).This kit is more sensitive than kit #7260 (figure 2).

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

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Companion Products

• 7150 PathScan® Total c-Jun Sandwich ELISA Kit II

Rabbit Monoclonals Produced UsingTechnology from Epitomics, Inc. UnderU.S. patent No. 5,675,063.