Measuring SHP-2 Phosphatase Activity

Immunoprecipitation and the measurement of SHP-2 activity were done as previously described [9 (link),23 (link)]. All the steps related to the SHP-2 activity were performed on ice and inside an anoxic chamber to prevent the spontaneous oxidation of the catalytic cysteine of SHP-2 (GB2202-P-V, MBraun, Garching Germany). The activity of SHP-2 was measured in the immunoprecipitates using the Malachite-Green based-Tyrosine Phosphatase Assay System (Promega, Walldorf, Germany).

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Dorenkamp M., Nasiry M., Semo D., Koch S., Löffler I., Wolf G., Reinecke H, & Godfrey R. (2023). Pharmacological Targeting of the RAGE-NFκB Signalling Axis Impedes Monocyte Activation under Diabetic Conditions through the Repression of SHP-2 Tyrosine Phosphatase Function. Cells, 12(3), 513.

Publication 2023

GB2202-P-V

Anoxic Assay Catalytic Cysteine Immunoprecipitation Malachite green Promega Shp 2 Tyrosine phosphatase

Corresponding Organization : University Hospital Münster

Other organizations : Jena University Hospital

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Variable analysis

independent variables

Immunoprecipitation and the measurement of SHP-2 activity

dependent variables

SHP-2 activity

control variables

All the steps related to the SHP-2 activity were performed on ice and inside an anoxic chamber to prevent the spontaneous oxidation of the catalytic cysteine of SHP-2

positive controls

None specified

negative controls

None specified

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