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PathScan® Phospho-c-Kit (Tyr719) Sandwich ELISA Kit

PathScan^® Phospho-c-Kit (Tyr719) Sandwich ELISA Kit #7298

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Supporting Data

REACTIVITY

Application Key:

ELISA-ELISA

Species Cross-Reactivity Key:

H-Human

Product Information

Product Description

CST's PathScan^® Phospho-c-Kit (Tyr719) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of c-Kit protein when phosphorylated at Tyr719. A c-Kit Mouse mAb has been coated onto the microwells. After incubation with cell lysates, both phospho- and nonphospho-c-Kit proteins are captured by the coated antibody. Following extensive washing, Phospho-c-Kit (Tyr719) Rabbit Antibody is added to detect the captured phospho-c-Kit protein. Anti-rabbit IgG HRP-linked Antibody #7074 is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of absorbance for this developed color is proportional to the quantity of c-Kit protein phosphorylated at Tyr719.

*Antibodies in kit are custom formulations specific to kit.

Protocol

Specificity / Sensitivity

CST's PathScan^® Phospho-c-Kit (Tyr719) Sandwich ELISA Kit #7298 detects endogenous levels of c-Kit protein when phosphorylated at Tyr719. A significant induction of phosphorylation of c-Kit at Tyr719 is detected in SCF-treated H526 cells using PathScan^® Phospho-c-Kit (Tyr719) Sandwich ELISA Kit #7298 (Figure 1). The level of total c-Kit protein in either nonphospho or phospho lysate detected by PathScan^® Total c-Kit Sandwich ELISA Kit #7197 or Western analysis remain unchanged (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.

Species Reactivity:

Human

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).

Database Links

UniProt ID: P10721

Entrez-Gene Id: 3815

限制使用

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

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PathScan is a registered trademark of Cell Signaling Technology, Inc.

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PathScan® Phospho-c-Kit (Tyr719) Sandwich ELISA Kit #7298