Product Pathways - PathScan ELISA
PathScan® Phospho-Stat3 (Tyr705) Sandwich ELISA Kit #7300
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|Kit Includes||Volume||Solution Color|
|Stat3 Monoclonal Ab Coated Microwells|
|P-Stat3 (Y705) Monoclonal Detection Ab||11 ml||Green|
|Anti-mouse IgG, HRP-linked Antibody||11 ml||Red|
|TMB Substrate #7004||11 ml||Colorless|
|STOP Solution #7002||11 ml||Colorless|
|Sealing Tape||2 sheets|
|ELISA Wash Buffer (20X)||25 ml||Colorless|
|ELISA Sample Diluent||25 ml|
|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).
H M R Mk
Species enclosed in parentheses are predicted to react based on 100% sequence homology.
CST's PathScan® Phospho-Stat3 (Tyr705) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Phospho-Stat3 (Tyr705) protein. A Stat3 rabbit monoclonal antibody has been coated onto the microwells. After incubation with cell lysates, both nonphospho- and phospho-Stat3 proteins are captured by the coated antibody. Following extensive washing, a Phospho-Stat3 (Tyr705) mouse monoclonal antibody is added to detect the captured phospho-Stat3 protein. HRP-linked anti-mouse antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of phospho-Stat3 protein.Antibodies in kit are custom formulations specific to kit.
Specificity / Sensitivity
CST's PathScan® Phospho-Stat3 (Tyr705) Sandwich ELISA Kit detects endogenous levels of Phospho-Stat3 protein. Using this Sandwich ELISA Kit #7300, a significant induction of phospho-Stat3 can be detected in IFN-α-treated HeLa cells. However, the level of total Stat3 protein remains unchanged, as shown by Western analysis (figure 1). This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
ELISA - Western correlation
Figure 1: Treatment of HeLa cells with IFN-alpha stimulates phosphorylation of Stat3 at Tyr705, detected by PathScan® Phospho-Stat3 (Tyr705) Sandwich ELISA kit, #7300, but does not affect the level of total Stat3 protein, detected by a Stat3 antibody via Western analysis. OD450 readings are shown in the top figure, while the corresponding Western blot using Phospho-Stat3 (Tyr705) (3E2) Monoclonal Antibody #9138 or Stat3 antibody, is shown in the bottom figure.
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).
- Heim, M.H. (2001) J Recept Signal Transduct Res 19, 75-120.
- Takeda, K. et al. (1997) Proc Natl Acad Sci U S A 94, 3801-4.
- Catlett-Falcone, R. et al. (1999) Immunity 10, 105-15.
- 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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- 7149 PathScan® Phospho-Stat3 (Tyr705) Chemiluminescent Sandwich ELISA Kit
- 7146 PathScan® Phospho-Stat3 (Tyr705) Sandwich ELISA Antibody Pair
- 7305 PathScan® Total Stat3 Sandwich ELISA Kit
- 9145 Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb
- 9131 Phospho-Stat3 (Tyr705) Antibody
- 4113 Phospho-Stat3 (Tyr705) (M9C6) Mouse mAb
- 9138 Phospho-Stat3 (Tyr705) (3E2) Mouse mAb
- 4904 Stat3 (79D7) Rabbit mAb
- 9139 Stat3 (124H6) Mouse mAb
- 7076 Anti-mouse IgG, HRP-linked Antibody
- 9803 Cell Lysis Buffer (10X)
- 7004 TMB Substrate
- 7002 STOP Solution
Rabbit Monoclonals Produced UsingTechnology from Epitomics, Inc. UnderU.S. patent No. 5,675,063.
For Research Use Only. Not For Use In Diagnostic Procedures.