Figure 1. Relationship between protein concentration of lysates from untreated and hEGF-treated A-431 cells and immediate light generation with chemiluminescent substrate is shown. Cells (85% confluence) were treated with hEGF #8916 (100 ng/ml) and lysed after incubation at 37ºC for 5 minutes. Graph inset corresponding to the shaded area shows high sensitivity and a linear response at the low protein concentration range.Learn more about how we get our images
|Product Includes||Volume||Solution Color|
|Stat3 Rabbit mAb Coated Microwells||96 tests|
The PathScan® Phospho-Stat3 (Ser727) Chemiluminescent Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-Stat3 (Ser727) 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 smaller samples. A Stat3 Rabbit mAb has been coated on the microwells. After incubation with cell lysates, Stat3 (phospho or nonphospho) protein is captured by the coated antibody. Following extensive washing, a Phospho-Stat3 (Ser727) Mouse mAb is added to detect the captured phospho-Stat3 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-Stat3 (Ser727) protein.
Antibodies in kit are custom formulations specific to kit.
PathScan® Phospho-Stat3 (Ser727) Chemiluminescent Sandwich ELISA Kit detects endogenous levels of phospho-Stat3 (Ser727) in human cells. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
The Stat3 transcription factor is an important signaling molecule for many cytokines and growth factor receptors (1) and is required for murine fetal development (2). Research studies have shown that Stat3 is constitutively activated in a number of human tumors (3,4) and possesses oncogenic potential (5) and anti-apoptotic activities (3). Stat3 is activated by phosphorylation at Tyr705, which induces dimerization, nuclear translocation, and DNA binding (6,7). Transcriptional activation seems to be regulated by phosphorylation at Ser727 through the MAPK or mTOR pathways (8,9). Stat3 isoform expression appears to reflect biological function as the relative expression levels of Stat3α (86 kDa) and Stat3β (79 kDa) depend on cell type, ligand exposure, or cell maturation stage (10). It is notable that Stat3β lacks the serine phosphorylation site within the carboxy-terminal transcriptional activation domain (8).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. PathScan is a trademark of Cell Signaling Technology, Inc.
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