WebMar 8, 2024 · APEX is a 27-kDa monomeric protein that promiscuously biotinylates proteins within a 20-nm radius of the target protein when supplied with ATP, H 2 O 2, and biotin-phenol at 37°C (Rhee et al., 2013).All neighboring proteins that are rich in tyrosine residues have the potential to be biotinylated by the APEX-fused bait protein within just 1 min, … WebJul 19, 2024 · Conversely, both control biotin tagging reactions via cytosolic APEX2 resulted in the enrichment of hundreds of cytosolic proteins or proteins facing the cytosolic side of organelles that were absent in MM …
An Optimized Protocol for Proximity Biotinylation in …
WebEnzymatic biotinylation with E. coli biotin ligase (BirA) is highly specific in covalently attaching biotin to the 15 amino acid AviTag peptide, giving a homogeneous product with high yield. AviTag can conveniently be added genetically at the N-terminus, C-terminus, or in exposed loops of a target protein. We describe here procedures for AviTag ... WebJan 29, 2024 · The APEX and HRP system. APEX is a monomeric 28 kDa ascorbate peroxidase that catalyzes the oxidative polymerization and local deposition of … shanghai academy of social sciences
APEX™ Antibody Labeling Kits - Thermo Fisher Scientific
WebSep 22, 2013 · By using a phenol derivative of biotin, APEX can thus catalyze proximity-based biotinylation of proteins. To demonstrate the use of APEX for compartment-specific biotinylation, APEX was targeted to the mitochondrial matrix and biotin–phenol and H 2 O 2 were added. MS analysis of purified biotinylated mitochondrial proteins identified several ... WebThe APEX™ Antibody Labeling Kits includes all reagents required to perform 5 separate labeling reactions of 10–20 µg of IgG antibody with one of Molecular Probes’ superior … WebBackground. NHS-Biotin (C 14 H 18 O 5 N 3 S) is N -hydroxysuccinimido biotin. It is the shortest of three similar EZ-Link NHS-Biotin Reagents that enable simple and efficient biotinylation of antibodies, proteins and any other primary amine-containing biomolecules in solution. NHS-Biotin offers researchers the possibility of optimizing labeling ... shanghai acceuil