Cell Signaling Technology

Product Pathways - PathScan ELISA

PathScan® Phospho-Stat3 (Tyr705) Sandwich ELISA Antibody Pair #7146

Kit Includes Volume Cap Color
Stat3 Capture Ab (100X) 0.4 ml Pink
P-Stat3 (Y705) Detection Ab (100X) 0.4 ml Blue
Anti-mouse IgG, HRP-linked Antibody (1000X) 0.04 ml Yellow

Capture and detection antibodies are stored at 4°C. HRP-linked secondary reagent is stored at -20°C.

Species Cross-Reactivity

H M

Reactivity Key:  H=Human  M=Mouse
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Description

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.

Specificity / Sensitivity

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.

Sandwich ELISA

Sandwich ELISA

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.

Background

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). 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).

  1. Heim, M.H. (2001) J Recept Signal Transduct Res 19, 75-120.
  2. Takeda, K. et al. (1997) Proc Natl Acad Sci U S A 94, 3801-4.
  3. Catlett-Falcone, R. et al. (1999) Immunity 10, 105-15.
  4. Garcia, R. and Jove, R. (1998) J Biomed Sci 5, 79-85.
  5. Bromberg, J.F. et al. (1999) Cell 98, 295-303.
  6. Darnell, J.E. et al. (1994) Science 264, 1415-21.
  7. Ihle, J.N. (1995) Nature 377, 591-4.
  8. Wen, Z. et al. (1995) Cell 82, 241-50.
  9. Yokogami, K. et al. (2000) Curr Biol 10, 47-50.
  10. Biethahn, S. et al. (1999) Exp Hematol 27, 885-94.

Application References

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Companion Products

Note: Antibody pairs have been optimized using recommended buffers, reagents, plates and protocol. Solutions should be made fresh daily.

For Research Use Only. Not For Use In Diagnostic Procedures.

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