Revision 4

#7300Store at +4C

1 个试剂盒

(96 assays)

Species Cross Reactivity

H M R Mk

UniProt ID：

#P40763

Entrez-Gene Id：

#6774

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Color Storage Temp
Stat3 Rabbit mAb Coated Microwells 40936 96 tests +4C
Phospho-Stat3 (Tyr705) Mouse Detection mAb 13149 1 ea Green (Lyophilized) +4C
Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated) 13304 1 ea Red (Lyophilized) +4C
Detection Antibody Diluent 13339 11 ml Green +4C
HRP Diluent 13515 11 ml Red +4C
TMB Substrate 7004 11 ml +4C
STOP Solution 7002 11 ml +4C
Sealing Tape 54503 2 ea +4C
ELISA Wash Buffer (20X) 9801 25 ml +4C
ELISA Sample Diluent 11083 25 ml Blue +4C
Cell Lysis Buffer (10X) 9803 15 ml -20C

*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.

Description

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.

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). 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).

  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.

Background References

    Cross-Reactivity Key

    H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    PathScan is a registered trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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    Orders: 877-616-CELL (2355)•[email protected]• Support: 877-678-TECH (8324)•[email protected]• Web: cellsignal.com
    仅供研究使用。不得用于诊断流程。
    Revision 4
    #7300

    PathScan® Phospho-Stat3 (Tyr705) Sandwich ELISA Kit

    PathScan® Phospho-Stat3 (Tyr705) Sandwich ELISA Kit: Image 1 Expand Image
    图 1：HeLa 细胞用 IFN-α 处理会刺激 Stat3 在 Tyr705 位点的磷酸化，使用 PathScan® Phospho-Stat3 (Tyr705) Sandwich ELISA kit #7300 可进行检测，但不影响使用 Stat3 antibody 通过蛋白质印迹分析检测到的 Stat3 总蛋白的水平。OD450 读数如顶部图所示，使用 Phospho-Stat3 (Tyr705) (3E2) Monoclonal Antibody #9138 或 Stat3 antibody 检测到的相应蛋白质印迹如底部图所示。
    PathScan® Phospho-Stat3 (Tyr705) Sandwich ELISA Kit: Image 2 Expand Image
    图 2：经 IFN-α 处理的 HeLa 细胞的裂解物蛋白浓度与使用试剂盒测得的光密度读数之间存在线性关系。HeLa 细胞（75% 融合度）用 IFN-α (100 ng/ml) 处理，并在 37o C 下生长 10 分钟后进行裂解。
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    Orders: 877-616-CELL (2355)•[email protected]• Support: 877-678-TECH (8324)•[email protected]• Web: cellsignal.com
    仅供研究使用。不得用于诊断流程。