Click-iT™ HPG Alexa Fluor™ 488 Protein Synthesis Assay Kit

Click-iT™ HPG Alexa Fluor™ 488 Protein Synthesis Assay KitCitations & References (5)Invitrogen™Click-iT™ HPG Alexa Fluor™ 488 Protein Synthesis Assay KitThe Click-iT™ HPG Alexa Fluor™ 488 Protein Synthesis Assay Kit provides a fast, sensitive, non-toxic, and non-radioactive method for theRead moreHave Questions?

Catalog Number	Quantity
C10428	1 kit

Catalog number C10428Price (USD) - Quantity:1 kitThe Click-iT™ HPG Alexa Fluor™ 488 Protein Synthesis Assay Kit provides a fast, sensitive, non-toxic, and non-radioactive method for the detection of nascent protein synthesis utilizing fluorescence microscopy, high-content imaging, or flow cytometry. Included in the kit are L-homopropargylglycine (HPG), an amino acid analog of methionine containing an alkyne moiety, and Alexa Fluor™ 488 azide. The HPG is fed to cultured cells and incorporated into proteins during active protein synthesis. Addition of the Alexa Fluor™ 488 azide leads to a chemoselective ligation or 'click' reaction between the green fluorescent azide and the alkyne, allowing the modified proteins to be detected by imaged-based analysis.• Non-radioactive alternative—an alternative to the traditional 35S-methionine • Visualize bulk protein dynamics—fluorescent tagging of proteins allows their localization to be determined, including aggregation• Specificity—selective, specific reaction between label and detection tags • Stability —product contains an irreversible, covalent bond • Multiplex-enabled—use in conjuction with Click™-iT AHA (azide amino acid and alkyne dye) to detect spatial and temporal differences • Applicability to biological samples—easy detection; high sensitivity and low background, regardless of complexityThe Click-iT™ HPG Alexa Fluor™ 488 Protein Synthesis Assay Kit has been successfully tested in HeLa, A549, and U-2 OS cells with a variety of reagents that inhibit protein synthesis, including cycloheximide and anisomycin. The applicability of these probes to monitor protein degradation has also been shown using inhibitors of the proteasome (MG132 and Bortezomib) and blockers of autophagy (chloroquine) in HeLa cells. Additionally, due to differences in Click-iT™ chemistry between the Click-iT™ HPG Alexa Fluor™ 488 Protein Synthesis Assay Kit and Click™-iT AHA with Alexa Fluor™ 594 Alkyne, these reagents can be used in conjunction for spatial or temporal determination of differences in nascent protein synthesis.For Research Use Only. Not for use in diagnostic procedures.SpecificationsDetection MethodFluorescenceDye TypeProteins (Nascent)FormLiquidGreen FeaturesLess hazardousQuantity1 kitColorGreenEmissionVisibleFor Use With (Application)Protein Synthesis AssayFor Use With (Equipment)Fluorescence Microscope, Fluorescent ImagerProduct LineAlexa Fluor, Click-iTProduct TypeProtein Synthesis Assay KitUnit SizeEachContents & Storage

• Store at 2°C to 6°C
• DO NOT FREEZE
• Protect material from long-term exposure to light, but may be exposed to light for short periods of time.

Citations & References (5)

Citations & ReferencesAbstractSelective identification of newly synthesized proteins in mammalian cells using bioorthogonal noncanonical amino acid tagging (BONCAT).Authors:Schuman EMJournal:Proceedings of the National Academy of Sciences of the United States of AmericaPubMed ID:16769897In both normal and pathological states, cells respond rapidly to environmental cues by synthesizing new proteins. The selective identification of a newly synthesized proteome has been hindered by the basic fact that all proteins, new and old, share the same pool of amino acids and thus are chemically indistinguishable. We ... MoreIn situ visualization and dynamics of newly synthesized proteins in rat hippocampal neurons.Authors:Schuman EMJournal:Nature neurosciencePubMed ID:20543841Protein translation has been implicated in different forms of synaptic plasticity, but direct in situ visualization of new proteins is limited to one or two proteins at a time. Here we describe a metabolic labeling approach based on incorporation of noncanonical amino acids into proteins followed by chemoselective fluorescence tagging ... MoreTwo-color labeling of temporally defined protein populations in mammalian cells.Authors:Tirrell DAJournal:Bioorganic & medicinal chemistry lettersPubMed ID:18774715The proteome undergoes complex changes in response to disease, drug treatment, and normal cellular signaling processes. Characterization of such changes requires methods for time-resolved protein identification and imaging. Here, we describe the application of two reactive methionine (Met) analogues, azidohomoalanine (Aha) and homopropargylglycine (Hpg), to label two protein populations in ... MoreFluorescence visualization of newly synthesized proteins in mammalian cells.Authors:Tirrell DAJournal:Angewandte Chemie (International ed. in English)PubMed ID:17036290Noncanonical amino acid tagging enables the selective fluorescent visualization of newly synthesized proteins in mammalian cells (see the picture). Susceptibility to tagging is determined by the spatial and temporal character of the protein synthesis, thus providing a complement to methods which identify relevant members of the proteome. ... MoreSpatial coupling of mTOR and autophagy augments secretory phenotypes.Authors:Narita M., et alJournal:Science (New York, N.Y.)PubMed ID:21512002Protein synthesis and autophagic degradation are regulated in an opposite manner by mammalian target of rapamycin (mTOR), whereas under certain conditions it would be beneficial if they occurred in unison to handle rapid protein turnover. We observed a distinct cellular compartment at the trans side of the Golgi apparatus, the ... More