Invitrogen™ Click-iT™ Plus OPP Alexa Fluor™ 647 Protein Synthesis Assay Kit

Description

• The Click-iT™ Plus OPP Alexa Fluor™ 647 Protein Synthesis Assay Kit provides a fast, sensitive, and non-radioactive method for the detection of protein synthesis using fluorescence microscopy or high-content imaging
• In this assay O-propargyl-puromycin (OPP) is efficiently incorporated into newly translated proteins in complete methionine-containing media and fluorescently labeled with a bright, photostable Alexa Fluor™ dye in a fast, highly-specific, and mild click reaction
• Features of the kit include: • No media change required—works in complete, methionine-containing media, no need to remove cell media • Multiplex-enabled—Click-iT™ Plus technology retains signal from GFP and binding of fluorescent-conjugated phalloidins • Non-radioactive—an alternative to the traditional 35 S-methionine methods • Works in vivo —published results demonstrate use in vivo for determination of protein translation The kit contains O-propargyl-puromycin (OPP), which is an alkyne analog of puromycin (also available separately), as well as Alexa Fluor™ 647 picolyl azide and all necessary reagents to perform the Click reaction
• The OPP is fed to cultured cells and incorporated into proteins during active protein synthesis
• Addition of the Alexa Fluor™ 647 picolyl azide and the Click reaction reagents leads to a chemoselective ligation, or 'click' reaction, between the picolyl azide dye and the OPP alkyne, allowing the modified proteins to be detected by imaged-based analysis
• The click reaction uses bioorthogonal (biologically unique) moieties to fluorescently label proliferating cells, helping to produce low backgrounds and high detection sensitivities
• Because of the mild reaction conditions, Click-iT™ Plus assays detect protein translation events while enabling preservation of cell morphology, the binding of fluorescently-labeled phalloidin, and the fluorescent signal from GFP
• Unlike 35 S-methionine, used in traditional methods, OPP is not an amino acid analog, so it can be added directly to cells in complete media or used to determine protein synthesis in vivo
• The kit contains all of the components needed to label and detect the incorporated OPP in newly translated proteins in samples of adherent cells
• The kit includes sufficient reagents for the labeling of 25 18 mm × 18 mm coverslips using 1 mL of reaction buffer per test.