SHP-2 (Full-Length) Homogeneous Assay Kit

Catalog #79330
$520
Size: 96 reactions

Description

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.

Materials Required But Not Supplied

Fluorescent microplate reader capable of reading exc/em=360 nm/455 nm
Adjustable micropipettor and sterile tips

Applications

Useful for studying enzyme kinetics and HTS applications.

Supplied As

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.

Stability

One year from date of receipt when stored as directed.

Background

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.

Synonym(s)

SHP2, CFC, NS1, BPTP3, PTP2C, PTP-1D, SH-PTP2, SH-PTP3, MGC14433, SH2, SH 2, SH-2, SHP, shp

Format

Catalog Number	Component	Amount	Storage
79018	Recombinant full length Human SHP-2	5 µg	-80°C	*Avoid freeze/ thaw cycles!*
79769	SHP-2 Substrate (1 mM)	50 µl	-80°C
79319-1	SHP-2 Activating Peptide (100 μM)	25 μl	-80°C
79626	5x SHP-2 Assay Buffer	3 ml	-20°C
	DTT (250 mM)	100 μl	-20°C
79685	Black, low binding black microtiter plate	1	Room Temp.

Instructions for Use

See assay kit data sheet for detailed protocol.

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

Notes

AlphaScreen^® and AlphaLISA^® are registered trademarks of PerkinElmer, Inc.

Scientific Category

Protein Phosphatase

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