The Homogeneous full length SHP-2 Assay Kit is a complete assay system designed to measure full length SHP-2 activity for screening and profiling applications. Using this kit, only two simple steps on a microtiter plate are needed to analyze the full length SHP-2 activity level. At the first step, the SHP-2 enzyme is preincubated with SHP-2 Activating Peptide to activate the enzyme. At the second step, the fluorogenic substrate, DiFMUP, is added in the mixture and the enzymatic activity releases DiFMU fluorophore that can then be measured using a fluorescence reader.
Kit comes in a convenient 96-well format, with all the reagents necessary for 100 fluorescent SHP-2 activity measurements. In addition, the kit includes purified full length SHP-2 enzyme for use as a positive control.
Materials Required But Not Supplied
Fluorescent microplate reader capable of reading exc/em=360 nm/460 nm Adjustable micropipettor and sterile tips
Mammalian PTPases can be subdivided into two broad categories: transmembrane receptor PTPases and intracellular PTPases. SHP-2 (PTPN11) is one of the 2 closely related mammalian intracellular PTPases whose sequences encode 2 tandem SRC homology 2 (SH2) domains located at the N-terminal side and a single PTPase catalytic domain at C-terminus. In the inactive state, the N-terminal SH2 domain interacts with the PTP catalytic domain and blocks access of potential substrates to the active site. Upon binding to target phospho-tyrosyl residues, the N-terminal SH2 domain is released from the PTP domain, and the enzyme is activated by relieving this autoinhibition. SHP-2 is widely expressed in most tissues and plays a regulatory role in various cell signaling events that are important for a diversity of cell functions, such as mitogenic activation, metabolic control, transcription regulation, and cell migration.
References
Chai, J., et al., Cell, 2001 Mar 9; 104(5):769-80. Denault, J.B., and Salvesen, GS., J. Biol. Chem. 2003 Sep 5; 278(36):34042-50. Fortanet, J.G., et al., J. Med. Chem., 2016, 59 (17):7773–7782
Storage/Stability
6 months from date of receipt when stored as directed.
Instructions for Use
See assay kit data sheet for detailed protocol.
Shipping Temperature
-80°C
Notes
AlphaScreen® and AlphaLISA® are registered trademarks of PerkinElmer, Inc.
Disclaimers and Limitations
BPS Bioscience assay kits are validated using the listed components according to our specific protocol. Any deviations to kit components or protocols, such as using different proteins, cell/tissue lysates, or buffers (including using the same component from other commercial sources) will void any warranties on the performance of the assay kit and is not recommended.
End-users should immediately report any issues to [email protected] upon receipt of the kit, such as missing components, damaged vials, no dry ice, etc.
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