PathScan® Phospho-Stat1 (Tyr701) Sandwich ELISA Kit #7234
Filter:
- ELISA
Supporting Data
| REACTIVITY | H |
Application Key:
- ELISA-ELISA
Species Cross-Reactivity Key:
- H-Human
Product Information
Product Description
The PathScan® Phospho-Stat1 (Tyr701) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Stat1 when phosphorylated at Tyr701. A Stat1 Rabbit Antibody has been coated onto the microwells. After incubation with cell lysates, Stat1 (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, biotinylated Phospho-Stat1 (Tyr701) Rabbit Detection Antibody is added to detect phosphorylation of Tyr701 on the captured Stat1 protein. HRP-linked streptavidin is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of Stat1 phosphorylated at Tyr701.
*Antibodies in kit are custom formulations specific to kit.
*Antibodies in kit are custom formulations specific to kit.
Protocol
Specificity / Sensitivity
CST's PathScan® Phospho-Stat1 (Tyr701) Sandwich ELISA Kit detects endogenous levels of Phospho-Stat1 when phosphorylated at Tyr701. As shown in Figure 1, a significant induction of Stat1 phosphorylation at Tyr701 can be detected in HeLa cells following treatment with Interferon-γ (IFN-γ) using the Phospho-Stat1 (Tyr701) Sandwich ELISA Kit #7234. The level of total Stat1 (phospho and nonphospho) remains unchanged as shown by Western analysis. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
Species Reactivity:
Human
Background
The Stat1 transcription factor is activated in response to a large number of ligands (1) and is essential for responsiveness to IFN-α and IFN-γ (2,3). Phosphorylation of Stat1 at Tyr701 induces Stat1 dimerization, nuclear translocation, and DNA binding (4). Stat1 protein exists as a pair of isoforms, Stat1α (91 kDa) and the splice variant Stat1β (84 kDa). In most cells, both isoforms are activated by IFN-α, but only Stat1α is activated by IFN-γ. The inappropriate activation of Stat1 occurs in many tumors (5). In addition to tyrosine phosphorylation, Stat1 is also phosphorylated at Ser727 through a p38 mitogen-activated protein kinase (MAPK)-dependent pathway in response to IFN-α and other cellular stresses (6). Serine phosphorylation may be required for the maximal induction of Stat1-mediated gene activation.
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