Product Pathways - PathScan ELISA
PathScan® Phospho-Stat3 (Ser727) Chemiluminescent Sandwich ELISA Kit #8026
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|Kit Includes||Volume||Solution Color|
|Stat3 Rabbit mAb Coated Microwells||96 tests|
|P-Stat3 (S727) Mouse Detection Antibody||5.5 ml||Green|
|Anti-mouse IgG, HRP-linked Antibody||5.5 ml||Red|
|Luminol/Enhancer Solution||3 ml||Colorless|
|Stable Peroxide Buffer||3 ml||Colorless|
|Sealing Tape||2 sheets|
|ELISA Wash Buffer (20X)||25 ml||Colorless|
|ELISA Sample Diluent||25 ml||Blue|
|Cell Lysis Buffer (10X) #9803||15 ml||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, which is stored at –20°C (packaged separately).
Species enclosed in parentheses are predicted to react based on 100% sequence homology.
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.
Specificity / Sensitivity
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.
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.
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).
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- Takeda, K. et al. (1997) Proc Natl Acad Sci U S A 94, 3801-4.
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- Garcia, R. and Jove, R. (1998) J Biomed Sci 5, 79-85.
- Bromberg, J.F. et al. (1999) Cell 98, 295-303.
- Darnell, J.E. et al. (1994) Science 264, 1415-21.
- Ihle, J.N. (1995) Nature 377, 591-4.
- Wen, Z. et al. (1995) Cell 82, 241-50.
- Yokogami, K. et al. (2000) Curr Biol 10, 47-50.
- Biethahn, S. et al. (1999) Exp Hematol 27, 885-94.
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* Product-specific protocol.
- 7995 PathScan® Phospho-Stat3 (Ser727) Sandwich ELISA Kit
- 7305 PathScan® Total Stat3 Sandwich ELISA Kit
- 7300 PathScan® Phospho-Stat3 (Tyr705) Sandwich ELISA Kit
- 7149 PathScan® Phospho-Stat3 (Tyr705) Chemiluminescent Sandwich ELISA Kit
- 7146 PathScan® Phospho-Stat3 (Tyr705) Sandwich ELISA Antibody Pair
- 9134 Phospho-Stat3 (Ser727) Antibody
- 9136 Phospho-Stat3 (Ser727) (6E4) Mouse mAb
- 7076 Anti-mouse IgG, HRP-linked Antibody
- 9803 Cell Lysis Buffer (10X)
For Research Use Only. Not For Use In Diagnostic Procedures.