The relationship between lysate protein concentration from untreated and IFN-α treated HeLa cells and the absorbance at 450 nm using PathScan® Phospho-Stat3 (Tyr705) Sandwich ELISA Antibody Pair #7146. After overnight starvation, HeLa cells were treated with IFN-α (100 ng/ml) for 10 minutes at 37ºC and then lysed.
|7146S||1 Kit (Reagents for 4 x 96 well plates)||$ 469|
|Product Includes||Volume||Cap Color|
|Stat3 Capture Rabbit mAb (100X)||400 µl||Pink|
|Phospho-Stat3 (Tyr705) Detection Mouse mAb (100X)||400 µl||Blue|
|Anti-mouse IgG, HRP-linked Antibody (1000X)||40 µl||Yellow|
Capture and detection antibodies are stored at 4°C. HRP-linked secondary reagent is stored at -20°C.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
1X Cell Lysis Buffer: 10X Cell Lysis Buffer (#9803): To prepare 10 ml of 1X Cell Lysis Buffer, add 1 ml of 10X Cell Lysis Buffer to 9 ml of dH2O, mix. Buffer can be stored at 4°C for short-term use (1–2 weeks).
Recommended: Add 1 mM phenylmethylsulfonyl fluoride (PMSF) (#8553) immediately before use.
STOP Solution: (#7002)
NOTE: Reagents should be made fresh daily.
posted January 2008
revised Sepetember 2013
Protocol Id: 20
CST's PathScan® Phospho-Stat3 (Tyr705) Sandwich ELISA Antibody Pair is offered as an economical alternative to our PathScan® Phospho-Stat3 (Tyr705) Sandwich ELISA Kit #7300. Capture and Detection antibodies (100X stock) and HRP-Linked Secondary Antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The Stat3 Rabbit Capture Antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added, followed by Phospho-Stat3 (Tyr705) Mouse Detection Antibody and HRP-Linked Anti-Mouse IgG. HRP substrate, TMB, is added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of phospho-Stat3 (Tyr705) protein.
Antibodies in kit are custom formulations specific to kit.
For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair 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).
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