Product Pathways - Tyrosine Kinase / Adaptors
PathScan® Phospho-Ret (panTyr) Chemiluminescent Sandwich ELISA Kit #12321
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|Kit Includes||Volume||Solution Color|
|Ret Rabbit mAb Coated Microwells||96 tests|
|Phospho-Tyrosine Mouse Detection mAb||5.5 ml||Green|
|Anti-mouse IgG, HRP-linked Antibody||5.5 ml||Red|
|Luminol/Enhancer Solution||3 ml||Colorless|
|Stable Peroxide Buffer||3 ml||Colorless|
|Sealing Tape||2 sheets|
|ELISA Wash Buffer (20X)||25 ml||Colorless|
|ELISA Sample Diluent||25 ml||Blue|
|Cell Lysis Buffer (10X) #9803||15 ml||Yellowish|
Note: 12 8-well modules –Each module is designed to break apart for 8 tests.
Note: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).
Species enclosed in parentheses are predicted to react based on 100% sequence homology.
The PathScan® Phospho-Ret (panTyr) Chemiluminescent Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of tyrosine-phosphorylated Ret protein with a chemiluminescent readout. Chemiluminescent ELISAs often have a wider dynamic range and higher sensitivity than conventional chromogenic detection. This chemiluminescent ELISA, which is offered in low volume microplates, shows increased signal and sensitivity while using a smaller sample size. A Ret Rabbit mAb has been coated onto the microwells. After incubation with cell lysates, Ret protein (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a Phospho-Tyrosine Mouse Detection mAb is added to detect captured tyrosine-phosphorylated Ret protein. Anti-mouse IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. Chemiluminescent reagent is added for signal development. The magnitude of light emission, measured in relative light units (RLU), is proportional to the quantity of tyrosine-phosphorylated Ret protein.Antibodies in kit are custom formulations specific to kit.
Specificity / Sensitivity
PathScan® Phospho-Ret (panTyr) Chemiluminescent Sandwich ELISA Kit #12321 detects endogenous levels of tyrosine-phosphorylated Ret protein in human cells. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
Figure 1. The relationship between protein concentration of lysates, untreated (Nonphospho) and phosphatase inhibitor treated (Phospho), and immediate light generation with chemiluminescent substrate is shown. TT cells were cultured (85% confluent) and lysed with the addition of phosphatase inhibitor to the PathScan® Sandwich ELISA Cell Lysis Buffer (1X) #9803. Graph inset corresponding to the shaded area shows high sensitivity and a linear response at the low protein concentration range.
The Ret proto-oncogene (c-Ret) is a receptor tyrosine kinase that functions as a multicompetent receptor complex in conjunction with other membrane-bound, ligand-binding GDNF family receptors (1). Ligands that bind the Ret receptor include the glial cell line-derived neurotropic factor (GDNF) and its congeners neurturin, persephin, and artemin (2-4). Research studies have shown that alterations in the corresponding RET gene are associated with diseases including papillary thyroid carcinoma, multiple endocrine neoplasia (type 2A and 2B), familial medullary thyroid carcinoma, and a congenital developmental disorder known as Hirschsprung’s disease (1,3). The Tyr905 residue located in the Ret kinase domain plays a crucial role in Ret catalytic and biological activity. Substitution of Phe for Tyr at position 905 dramatically inhibits Ret autophosphorylation activity (5).
- Airaksinen, M.S. et al. (1999) Mol. Cell. Neurosci. 13, 313-325.
- Takahashi, M. et al. (1989) Oncogene 4, 805-806.
- Manie, S. et al. (2001) Trends Genet. 17, 580-589.
- Tallini, G. and Asa, S. (2001) Adv. Anat. Pathol. 8, 345-354.
- Iwashita, T. et al. (1999) Oncogene 18, 3919-3922.
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* Product-specific protocol.
- 7034 PathScan® Phospho-Ret (panTyr) Sandwich ELISA Kit
- 7032 PathScan® Total Ret Sandwich ELISA Kit
- 9803 Cell Lysis Buffer (10X)
For Research Use Only. Not For Use In Diagnostic Procedures.