Phosphotyrosine c-Kit ELISA Kit is a Semi-quantitative ELISA kit for the measurement of Phosphotyrosine c-Kit in Human in Cell Lysate samples.
Application | Reactivity | Dilution info | Notes |
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Application ELISA | Reactivity Reacts | Dilution info - | Notes - |
Phosphotyrosine c-Kit ELISA Kit is a Semi-quantitative ELISA kit for the measurement of Phosphotyrosine c-Kit in Human in Cell Lysate samples.
Phosphotyrosine c-Kit ELISA Kit (ab279918) is a very rapid, convenient, and sensitive assay kit that can monitor the activation or function of important biological pathways in human cell lysates. By determining phosphorylated c-Kit protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.
This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human phospho-c-Kit. An anti-c-Kit antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and phosphorylated and unphosphorylated c-Kit present in a sample is bound to the wells by the immobilized antibody. The wells are washed, and biotinylated anti-phosphotyrosine antibody is used to detect only tyrosine-phosphorylated protein. After washing away unbound antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of phospho-c-Kit bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
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The c-Kit protein also known as CD117 is a type of receptor tyrosine kinase with a molecular weight of approximately 145 kDa. It is encoded by the KIT gene and commonly expressed on the surface of hematopoietic stem cells mast cells melanocytes and germ cells. C-Kit plays a role in the signal transduction pathways that control cell proliferation and survival. Researchers often identify c-Kit using markers like biotin c-Kit PerCP Cy5.5 or antibodies such as anti-c-Kit for experimental purposes.
C-Kit influences the function and survival of certain cell types. When it binds to its ligand stem cell factor (SCF) it activates receptor dimerization and phosphorylation triggering downstream signaling. C-Kit does not act alone; it often functions in a complex with other proteins to affect cellular outcomes. Its expression is vital for the maintenance of bone marrow hematopoietic cells and contributes to melanocyte and germ cell development.
C-Kit interacts with the PI3K/AKT and RAS/ERK pathways through its kinase activity. These pathways play important roles in cell division differentiation and survival. The c-Abl and ACK2 proteins are closely related to c-Kit function within these signaling networks contributing to regulating cellular responses to external stimuli.
C-Kit mutations are frequently associated with gastrointestinal stromal tumors (GIST) and acute myeloid leukemia (AML). These conditions often involve alterations in c-Kit activity leading to uncontrolled cell growth. While studying GIST researchers have noticed interactions between c-Kit and other proteins like CBL which is involved in the modulation of c-Kit signaling. Understanding these interactions helps in designing targeted therapies for conditions involving c-Kit dysregulation.
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Positive Control.
Jurkat cells were treated with Pervanadate at 37°C for 10 min.
Solubilize cells at 4 x 107 cells/ml in lysis buffer.
Serial dilutions of lysates were analyzed in this ELISA.
HeLa cells treated/untreated with pervanadate.
HeLa cells were untreated or treated with Pervanadate for 10 miutesn at 37°C.
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