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Citations & References (19)
Invitrogen™
Click-iT™ AHA Alexa Fluor™ 488 Protein Synthesis HCS Assay
The Click-iT® AHA Alexa Fluor® 488 Protein Synthesis HCS Assay provides a fast, sensitive, non-toxic and non-radioactive method for theRead more
| Catalog Number | Quantity |
|---|---|
| C10289 | 2 plates |
Catalog number C10289
Price (CLP)
920.944
Each
Quantity:
2 plates
Price (CLP)
920.944
Each
The Click-iT® AHA Alexa Fluor® 488 Protein Synthesis HCS Assay provides a fast, sensitive, non-toxic and non-radioactive method for the detection of nascent protein synthesis utilizing fluorescence microscopy and high-content imaging. Click-iT® AHA is an amino acid analog of methionine containing an azido moiety. Similar to 35S-methionine, Click-iT® AHA is fed to cultured cells and incorporated into proteins during active protein synthesis. Detection of the incorporated amine acid utilizes a chemoselectiove ligation or “click" reaction between an azide and an alkyne, where the azido modified protein is detected with the green-fluorescent Alexa Fluor® 488 alkyne. The Click-iT® AHA Alexa Fluor® 488 Protein Synthesis HCS Assay has been successfully tested in A549 and U-2 OS cells with a variety of reagents that inhibit protein synthesis including cycloheximide, anisomycin and puromycin in both dose response and Min⁄Max format.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Detection MethodFluorescence
Dye TypeAlexa Fluor™ 488, Hoechst 33258
Format96-well plate
Green FeaturesLess hazardous
Quantity2 plates
Shipping ConditionRoom Temperature
ColorBlue, Green
For Use With (Application)High-content Screening (HCS)
For Use With (Equipment)Fluorescence Microscope, High Content Analysis Instrument
Product LineAlexa Fluor, Click-iT, Molecular Probes
Product TypeHCS Assay
Unit SizeEach
Contents & Storage
The kit contains sufficient reagents for performing 2 plates in a 96-well plate format.Store at ≤-20°C, desiccate and protect from light.
Frequently asked questions (FAQs)
I am using Click-iT AHA (L-azidohomoalanine) kit to label nascent proteins in live cells, then detecting with TAMRA alkyne after fixation and permeabilization and a click reaction. But I'm seeing nucleolar labeling in the cells. It this expected?
Citations & References (19)
Citations & References
Abstract
Selective identification of newly synthesized proteins in mammalian cells using bioorthogonal noncanonical amino acid tagging (BONCAT).
Journal:Proceedings of the National Academy of Sciences of the United States of America
PubMed ID:16769897
In 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
In situ visualization and dynamics of newly synthesized proteins in rat hippocampal neurons.
Journal:Nature neuroscience
PubMed ID:20543841
Protein 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
Two-color labeling of temporally defined protein populations in mammalian cells.
Journal:Bioorganic & medicinal chemistry letters
PubMed ID:18774715
The 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
Fluorescence visualization of newly synthesized proteins in mammalian cells.
Journal:Angewandte Chemie (International ed. in English)
PubMed ID:17036290
Noncanonical 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.
Spatial coupling of mTOR and autophagy augments secretory phenotypes.
Journal:Science (New York, N.Y.)
PubMed ID:21512002
Protein 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