Basic Information
ID: ALA3705485
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.
Organism: Homo sapiens
Activity Charts
Bioactivity
Activity Types for Assay ALA3705485
IC50
Compound Summary
| Parent Molecular Weight | ALogP | Polar Surface Area |
|---|---|---|
| 320.73 | 2.01 | 2.01 |
| 320.73 | 2.01 | 2.01 |
| 311.30 | 1.91 | 1.91 |
| 286.29 | 1.58 | 1.58 |
| 285.30 | 2.18 | 2.18 |
| 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 |
| 314.34 | 2.39 | 2.39 |
| 290.25 | 1.41 | 1.41 |
| 330.34 | 2.06 | 2.06 |
| 320.73 | 2.23 | 2.23 |
| 319.75 | 1.58 | 1.58 |
| 311.30 | 1.91 | 1.91 |
| 354.33 | 0.54 | 0.54 |
| 297.27 | 1.82 | 1.82 |
| 329.31 | 0.46 | 0.46 |
| 383.40 | 1.59 | 1.59 |
| 369.38 | 1.50 | 1.50 |
| 340.30 | 0.45 | 0.45 |
| 369.38 | 1.71 | 1.71 |