ID: ALA3707537
Type: Binding
Description: Mass Spectrometry Assay : The EGLN-1 (or EGLN-3) enzyme activity is determined using mass spectrometry (matrix-assisted laser desorption ionization, time-of-flight MS, MALDI-TOF MS—for assay details, see reference (Greis et al, 2006). The reaction is conducted in a total volume of 50 uL containing TrisCl (5 mM, pH7.5), ascorbate (120 μM), 2-oxoglutarate (3.2 μM), HIF-1α (8.6 μM), and bovine serum albumin (0.01%). The enzyme, quantity predetermined to hydroxylate 20% of substrate in 20 minutes, is added to start the reaction. Compounds are prepared in dimethyl sulfoxide at 10-fold final assay concentration. After 20 minutes at room temperature, the reaction is stopped by transferring 10 μL of reaction mixture to 50 μL of a mass spectrometry matrix solution (α-cyano-4-hydroxycinnamic acid, 5 mg/mL in 50% acetonitrile/0.1% TFA, 5 mM NH4PO4). Two microliters of the mixture is spotted onto a MALDI-TOF MS target plate for analysis with an Applied Biosystems.
Format: BAO_0000357
Organism: Homo sapiens
Target: Egl nine homolog 3(ALA5705)
Document: ALA3638372
BindingDB Source: 6302
Bioactivity
Activity Types for Assay ALA3707537
IC50
Parent Molecular Weight | ALogP | Polar Surface Area |
---|---|---|
320.73 | 2.01 | 2.01 |
286.29 | 1.58 | 1.58 |
300.31 | 1.67 | 1.67 |
320.73 | 2.01 | 2.01 |
297.27 | 1.82 | 1.82 |
306.71 | 1.92 | 1.92 |
306.71 | 1.92 | 1.92 |
306.71 | 1.92 | 1.92 |
319.75 | 1.58 | 1.58 |