Revision 6

#7145

Store at +4C

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

1 Kit

(96 assays)

Species Cross Reactivity:
H M R

UniProt ID:
#P05412

Entrez-Gene Id:
#3725

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Product Includes Product # Quantity Color Storage Temp
Phospho-c-Jun (Ser63) RmAb Coated Microwells2902796 tests+4C
c-Jun Mouse Detection mAb142571 eaGreen (Lyophilized)+4C
Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated)133041 eaRed (Lyophilized)+4C
Detection Antibody Diluent1333911 mlGreen+4C
HRP Diluent1351511 mlRed+4C
TMB Substrate700411 ml+4C
STOP Solution700211 ml+4C
Sealing Tape545032 ea+4C
ELISA Wash Buffer (20X)980125 ml+4C
ELISA Sample Diluent1108325 mlBlue+4C
Cell Lysis Buffer (10X)980315 ml-20C

Kit contents scale proportionally with size, except sealing tape.
Example: The V1 kit contains 5X the listed quantities above, but will exclude the sealing tape.

For the “C” and “V” kits, the supplied 96-well strip plate consists of twelve 8-well strips in a support frame. This enables custom plate configurations.

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 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 is added to detect the captured phospho-c-Jun protein. Anti-Mouse IgG, HRP-linked antibody 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 kit are custom formulations specific to 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 a Total c-Jun Sandwich ELISA, remains unchanged. Both C6 and NIH/3T3 cells treated with UV light or anisomycin show similar results (data not shown). This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Background

c-Jun is a member of the Jun family containing c-Jun, JunB, and JunD, and is a component of the transcription factor activator protein-1 (AP-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). Knockout 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). Research studies have implicated c-Jun as a promising therapeutic target for cancer, vascular remodeling, acute inflammation, and rheumatoid arthritis (11,12).

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

PathScan is a registered trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

Cell Signaling Technology Logo
Figure 1. Treatment of 293 cells with UV stimulates phosphorylation of c-Jun at Ser63, but does not affect the level of total c-Jun. The relationship between lysate protein concentration from untreated and UV treated 293 cells and the absorbance at 450 nm using the PathScan® Phospho-c-Jun (Ser63) Sandwich ELISA Kit II is shown in the upper figure. The corresponding western blots using phospho-cJun (Ser63) antibody are shown in the lower figure. Lambda protein phosphatase (LPP) treatment of control cell lysates (37°C for 90 minutes) abolished the basal phosphorylation of c-Jun as shown in Western analysis. 293 cells (70-90% confluence) were treated with or without UV and lysed after incubation at 37°C for 30 minutes.
ELISA Image 1: PathScan<sup>®</sup> Phospho-c-Jun (Ser63) Sandwich ELISA Kit II
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.