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c-Jun Phospho (pT91/93) ELISA Kit antibody RabMAb®

Cat.#: 6115-1

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A Simple, Complete ELISA Kit

All-In-One: Quick and easy sandwich ELISA Kit
Sensitive: Useful for detection of levels of c-Jun Phospho (pT91/93)

Description

c-Jun is a transcription factor that recognizes and binds to the enhancer heptamer motif 5-TGA [CG] TCA-3. c-Jun forms homodimers and heterodimers with Fos and other jun-related proteins which, together, comprise the AP-1 transcription factor that binds TPA response elements (TREs). c-Jun therefore mediates transcriptional regulation in response to a variety of stimulants (1). c-Jun is tightly regulated posttranslationally and is phosphorylated in two distinct regions. Ser 63 and 73 are required to be phosphorylated for efficient transactivation function. The kinases responsible for this modification in vivo are the SAPK/JNKs (2, 3). Transactivation of c-Jun is regulated by Jun-N-terminal kinases (JNKs) through phosphorylation at serine 63 and 73 (S63/S73), as well as at threonine 91 and 93 (T91/T93). The integrity of JNK phosphorylation sites at serines 63/73 and at threonines 91/93 in c-Jun is essential for signal-dependent target gene activation (4). These two groups of phosphoacceptor sites respond to different grades of genotoxic stress. c-Jun phosphorylation is restricted to S63/S73, following a short exposure to the DNA-damaging compound etoposide. In contrast, JNK-dependent phosphorylation of T91/T93 requires continuous exposure to the drug (5). This assay employs the sandwich enzyme immunoassay technique that detects endogenous level of c-Jun when phosphorylated at Threonine 91 and 93. Rabbit anti-c-Jun monoclonal antibody has been precoated onto the microplate. Cell lysate is pipetted into the wells. Following extensive washing, biotinylated rabbit anti-c-Jun Phospho (pT91/93) monoclonal antibody reagent is added. Following a wash to remove any unbound antibody reagent, streptavidin-HRP is added to the well. After washing away the unbound streptavidin-HRP, a substrate solution is added to the wells to develop color. The magnitude of the absorbance for this developed color is proportional to the amount of endogenous phosphorylated c-Jun.

Recommended Applications

ELISA

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Products Data

Fig 1. Main steps of the c-Jun Phospho (pT91/93) Sandwich ELISA assay.	Fig 1. Treatment of A431 cells with TSA (500 ng/ml for 4 hr) increases the acetylation of Histone.	Fig 2. The relationship between the protein concentration of lysates from untreated and TSA-treated A431 cells and assay optical density readings.

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Components

Dry Plate with stripwells (96)
Biotinylated c-Jun Phospho (pT91/93) Antibody Reagent
Streptavidin-HRP Reagent
Antigen/Antibody Diluent Buffer
ELISA Washing Buffer
TMB A Substrate Solution
TMB B Substrate Solution
Stop Solution

Storage Condition and Buffer

Store at -20 °C. Buffer: 50 mM Tris-Glycine (pH 7.4), 0.15 M NaCl, 40% Glycerol, 0.01% sodium azide and 0.05% BSA. Stable for 12 months from date of receipt.

Description References

1. Franklin, C.C., Sanchez, V., Wagner, F., Woodgett, J.R. and Kraft, A.S. (1992) Proc. Natl. Acad. Sci. USA 89, 7247-7251 2. Kallunki, T., Su, B., Tsigelny, I., Sluss, H.K., Derijard, B., Moore, G., Davis, R. and Karin, M. (1994) Genes and Development 8, 2996-3007 3. Pulverer, B., Kyriakis, J., Avruch, J., Nikolakaki, H. and Woodgett, J.R. (1991) Nature 353, 670-674 4. Weiss C, Schneider S, Wagner EF, Zhang X, Seto E, Bohmann D. (2003) EMBO J. 22(14):3686-95 5. Vinciguerra M, Esposito I, Salzano S, Madeo A, Nagel G, Maggiolini M, Gallo A, Musti AM. (2008) Int J Biochem Cell Biol. 40(2):307-16.

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