Inhibition of HO-1 (unknown origin) expressed in human imatinib mesylate-resistant LAMA-84 cells assessed as bilirubin formation at 10 uM after 24 hrs relative to control
Kinobeads (epsilon), multiple immobilized ATP-competitive broad spectrum kinase inhibitors, used to assess residual binding of ~300 proteins simultaneously from cell lysate in the presence of a compound. Quantitative readout performed by mass spectrometry.
Binding affinity to HMOX2 cysteine residue in human 786-O cell soluble proteomic lysate at 5 uM incubated for 1 hr followed by cell lysis by IA-alkyne probe based isoTOP-ABPP analysis
Binding affinity to HMOX2 cysteine residue in human 786-O cell lysate at 5 uM incubated for 1 hr followed by cell lysis by IA-alkyne probe based isoTOP-ABPP analysis
Binding affinity to HMOX2 in human A549 cells lysates grown on SILAC media at 10 uM incubated for 1 hr by LC-MS/MS analysis relative to untreated control
Binding affinity to HMOX2 in human HePAD38 cells infected with HBV assessed as protein labelling at 20 uM after 1 hr by photoaffinity labelling in-situ fluorescence gel-based LC-MS/MS analysis
Covalent binding affinity to HMOX2 at Cys282 residue in human NCI-H358 cells assessed as log2 H/L ratio at 10 uM pretreated for 3 hrs followed by N-(((6S,7R)-7-(4-chloro-3-fluorophenyl)-1,4-oxazepan-6-yl)methyl)-2-hydroxyacetamide treatment for 3 hrs by competitive TMT click chemistry pull-down experiment based LC-MS/MS analysis
Covalent binding affinity to HMOX2 at Cys282 residue in human NCI-H358 cells assessed as reduction of log2 H/L ratio for tryptic peptide GALEGSSCPFR incubated for 4 hrs followed by cell lysis and subsequently labelled with light thiol probe N-((S)-18-iodo-11-isopropyl-10,13,17-trioxo-3,6-dioxa-9,12,16-triazaoctadecyl)-6-((4R,5S)-5-methyl-2-oxoimidazolidin-4-yl)hexanamide and heavy thiol probe 4 for 1 hr under dark conditions followed by tryptic digestion for overnight and measured by discovery ABPP based nano LC-MS/MS analysis