The PathScan® Phospho-M-CSF Receptor (Tyr699) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of M-CSF receptor protein phosphorylated at Tyr699. A M-CSF receptor mouse mAb has been coated onto the microwells. After incubation with cell lysates, both phospho- and nonphospho-M-CSF receptor proteins are captured by the coated antibody. Following extensive washing, Phospho-M-CSF Receptor (Tyr699) Rabbit Detection mAb is added to detect the captured phospho-M-CSF receptor proteins. Anti-rabbit IgG, HRP-linked 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 M-CSF receptor protein phosphorylated at Tyr699.
Antibodies in kit are custom formulations specific to kit.
PathScan® Phospho-M-CSF Receptor (Tyr699) Sandwich ELISA Kit detects endogenous levels of M-CSF receptor protein phosphorylated at Tyr699 in human cells, as shown in Figure 1. The kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
Macrophage-colony stimulating factor (M-CSF, CSF-1) receptor is an integral membrane tyrosine kinase encoded by the c-fms proto-oncogene. M-CSF receptor is expressed in monocytes (macrophages and their progenitors) and drives growth and development of this blood cell lineage. (1-3). Binding of M-CSF to its receptor induces receptor dimerization, activation, and autophosphorylation of cytoplasmic tyrosine residues used as docking sites for SH2-containing signaling proteins (4). There are at least five major tyrosine autophosphorylation sites. Tyr723 (Tyr721 in mouse) is located in the kinase insert (KI) region. Phosphorylated Tyr723 binds the p85 subunit of PI3 kinase as well as PLCγ2 (5). Phosphorylation of Tyr809 provides a docking site for Shc (5). Overactivation of this receptor can lead to a malignant phenotype in various cell systems (6). The activated M-CSF receptor has been shown to be a predictor of poor outcome in advanced epithelial ovarian carcinoma (7) and breast cancer (8).
Phosphorylation of M-CSF receptor at Tyr699 was identified at Cell Signaling Technology (CST) using PhosphoScan®, CST's LC-MS/MS platform for phosphorylation site discovery, and was cited in another publication (10). Autophosphorylation at Tyr699 within the kinase insert (KI) domain appears to provide a binding site for the Grb2 adaptor protein (9).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. PathScan is a trademark of Cell Signaling Technology, Inc.
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