MILPITAS, Calif. — Palmitoylation occurs when fatty acids, such as Palmitic Acid are covalently attached to the side chains of cysteine (and less frequently to serine, threonine side chains) in proteins. This type of post-translational modification greatly affects cellular trafficking, compartmentalization, and membrane tethering. Palmityltransferase (PAT), the enzyme responsible for this modification transfers a palmitate moiety from palmitoyl-CoA to the thiol group of cysteine in the target protein. Compared to myristoylation, palmitoylation is reversible and the reverse reaction is catalyzed by thioesterases. The Palmitoylation/Depalmitoylation cycle plays an important role when modified protein shuttles between cellular compartments. PAT mutations are associated with many neurological diseases and cancer progression.

BioVision offers EZClickTM Palmitoylated Protein Assay Kit, a highly specific, simple and robust method for labeling and detection of palmitoylated proteins. The kit uses a modified Palmitic Acid that is fed directly into the cells and gets incorporated into proteins during or post translation. This post translational modification can be followed by click reaction with an azide-containing dye. The assay kit offers a powerful method for imaging localization, trafficking, and dynamics of Palmitoylated proteins, or detection by FACS for quantitative studies. Labeled Palmitoylated Proteins can be directly detected in 1D or 2D gels using the appropriate excitation sources, or enriched by immunoprecipitation with biotin-azide or antibodies prior to proteomic analysis. We provide sufficient materials for 100 assays in a 96-well plate format.

Figure:Jurkat cells: FACS analysis of Negative Control (Black), Background (EZClick only, Green), Positive Control (Palmitic Acid Label and EZClick, Pink). Signal measured in FL-1 channel clearly shows the Palmitic Acid Labeling of protein.  Assays were performed using kit K452-100 protocol.

For more information on this kit, visit: https://www.biovision.com/ezclicktm-palmitoylated-protein-assay-kit-facs-microscopy-green-fluorescence.html