| Assay Method Information | |
| | recombinant human MetAP2 assay |
| Description: | The plate was sealed with a TopSeal A cover and mixed briefly on an orbital mixer at 900 rpm. The plate was incubated for a further 25 minutes at 25 ° C. A 5 stock of Amplex buffer was prepared =(0.25M sodium phosphate, pH 7.4) and stored at 4 ° C. When preparing for use the stock was diluted with distilled water. Amplex Ultraread stock solution was prepared at 2.57 mg/ml in 100% DMSO and stored in 50 l aliquots at 20 ° C. 20 l of 505 U/ml. Horse radish peroxidase was diluted in 990 ml of Amplex buffer, 100 l of this was combined with 50 l of Amplex Ultrared in 4850 ml of 1 Amplex buffer to generate sufficient detection reagent for a 384 well plate. 25 l detection reagent was added to each well of the test plate, which was re-sealed and mixed briefly on an orbital shaker. The plate was transferred to an Envision Multi-label reader and RFU measured corresponding to excitation 531 nm and emission 595 nm. At the end of the MetAP2 incubation 25 l Amplex/HRP mixture per well was added and the plate read plate on a plate reader. |
| Affinity data for this assay | |
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