Relationship between protein concentration of lysates from untreated and UV-treated 293 cells and immediate light generation with chemiluminescent substrate is shown. 293 cells (70-90% confluence) were treated with or without UV and lysed after incubation at 37ºC for 30 minutes. The graph inset corresponding to the shaded area shows high sensitivity and a linear response at the low protein concentration range.
|REACTIVITY||H M R|
|Product Includes||Volume||Solution Color|
|Phospho-c-Jun (Ser63) Rabbit mAb Coated Microwells||96 tests|
|c-Jun Mouse Detection mAb||1 ea||Green (Lyophilized)|
|Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated)||1 ea||Red (Lyophilized)|
|Detection Antibody Diluent||5.5 ml||Green|
|HRP Diluent||5.5 ml||Red|
|Luminol/Enhancer Solution||3 ml|
|Stable Peroxide Buffer||3 ml|
|Sealing Tape||2 ea|
|ELISA Wash Buffer (20X) 9801||25 ml|
|ELISA Sample Diluent||25 ml||Blue|
|Cell Lysis Buffer (10X) 9803||15 ml|
The PathScan® Phospho-c-Jun (Ser63) Chemiluminescent Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-c-Jun (Ser63) protein with a chemiluminescent readout. Chemiluminescent ELISAs often have a wider dynamic range and higher sensitivity than conventional chromogenic detection. This chemiluminescent ELISA, which is offered in low volume microplates, shows increased signal and sensitivity while using a smaller sample size. A phospho-c-Jun (Ser63) rabbit mAb has been coated on the microwells. After incubation with cell lysates, phospho-c-Jun 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. Chemiluminescent reagent is added for signal development. The magnitude of light emission, measured in relative light units (RLU), is proportional to the quantity of phospho-c-Jun (Ser63) protein.
Antibodies in kit are custom formulations specific to kit.
NOTE: Refer to product-specific datasheets for assay incubation temperature. This chemiluminescent ELISA is offered in low volume microplates. Only 50 µl of samples or reagents are required in each microwell.
NOTE: Prepare solutions with purified water.
*NOTE: Some PathScan® ELISA Kits may include HRP-Linked Streptavidin in place of HRP-Linked Antibody.
posted November 2013
Protocol Id: 66
PathScan® Phospho-c-Jun (Ser63) Chemiluminescent Sandwich ELISA Kit #7027 detects endogenous levels of phospho-c-Jun (Ser63) in human, mouse and rat cells. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
Human, Mouse, Rat
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). Knock-out 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).
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