| Product Includes | Product # | Quantity | Color | Storage Temp |
|---|---|---|---|---|
| Phospho Tyrosine Mouse Detection mAb (Biotinylated) | 13770 | 1 ea |
|
+4C |
| HRP-Linked Streptavidin (ELISA Formulated) | 11805 | 1 ea |
|
+4C |
| Detection Antibody Diluent | 13339 | 11 ml |
|
+4C |
| HRP Diluent | 13515 | 11 ml |
|
+4C |
| TMB Substrate | 7004 | 11 ml |
|
+4C |
| STOP Solution | 7002 | 11 ml |
|
+4C |
| ELISA Wash Buffer (20X) | 9801 | 25 ml |
|
+4C |
| ELISA Sample Diluent | 11083 | 25 ml |
|
+4C |
| Cell Lysis Buffer (10X) | 9803 | 15 ml |
|
-20C |
| Sealing Tape | 54503 | 2 ea |
|
+4C |
*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.
Description
The PathScan® Phospho-c-Kit (panTyr) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of c-Kit when tyrosine phosphorylated. A c-Kit Mouse mAb has been coated onto the microwells. After incubation with cell lysates, c-Kit (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a Biotinylated Phospho-Tyrosine Mouse Detection Antibody is added to detect tyrosine phosphorylation of the captured c-Kit protein. HRP-linked Strepavidin 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 c-Kit phosphorylated on tyrosine.
Antibodies in kit are custom formulations specific to kit.
Specificity/Sensitivity
Background
c-Kit is a member of the subfamily of receptor tyrosine kinases that includes PDGF, CSF-1, and FLT3/flk-2 receptors (1,2). It plays a critical role in activation and growth in a number of cell types, including hematopoietic stem cells, mast cells, melanocytes, and germ cells (3). Upon binding with its stem cell factor (SCF) ligand, c-Kit undergoes dimerization/oligomerization and autophosphorylation. Activation of c-Kit results in the recruitment and tyrosine phosphorylation of downstream SH2-containing signaling components, including PLCγ, the p85 subunit of PI3 kinase, SHP2, and CrkL (4). Molecular lesions that impair the kinase activity of c-Kit are associated with a variety of developmental disorders (5), and mutations that constitutively activate c-Kit can lead to pathogenesis of mastocytosis and gastrointestinal stromal tumors (6). Tyr719 is located in the kinase insert region of the catalytic domain. c-Kit phosphorylated at Tyr719 binds to the p85 subunit of PI3 kinase in vitro and in vivo (7).
- Martin, F.H. et al. (1990) Cell 63, 203-11.
- Yarden, Y. et al. (1987) EMBO J 6, 3341-51.
- Gommerman, J.L. et al. (1997) J Biol Chem 272, 30519-25.
- Sattler, M. et al. (1997) J Biol Chem 272, 10248-53.
- Nocka, K. et al. (1990) EMBO J 9, 1805-13.
- Hirota, S. et al. (1998) Science 279, 577-80.
- Blume-Jensen, P. et al. (2000) Nat Genet 24, 157-62.
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
限制使用
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