| Assay Method Information | |
| | Assay for the inhibitory effect on the SHP2 enzyme |
| Description: | A mixed liquor of 2× working SHP2 reagent (0.4 nM) and 0.5 μM SHP-2 activated peptide (BPS bioscience #79319-2) was prepared with 1× enzyme buffer. After the above-mixed liquor was incubated at 25° C. for 60 minutes, 5 μL of activated SHP2 kinase solution was transferred with a pipettor to the compound wells and the Max control wells (Corning, #4514) of the plate. Then 5 μL of 1× kinase buffer was transferred to the Min control wells. The plate was centrifuged at 1000 rpm for 30 seconds, sealed, and incubated in a 25° C. constant-temperature incubator for 30 minutes. A substrate solution of DiFMUP (#D6567, Invitrogen™) was prepared with 1× kinase buffer, and its concentration should be 2 times as the final concentration of the assay (DiFMUP final concentration: 10 μM). 5 μL of the prepared substrate solution was transferred with an electric pipettor into each well of the ELISA plate to start the reaction. The plate was centrifuged at 1000 rpm for 30 seconds, sealed, and incubated in a 25° C. constant-temperature incubator for 60 minutes. Read the plate at the excitation/emission wavelength of 358/455 nm after the ELISA plate was placed on the Spark machine. |
| Affinity data for this assay | |
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