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Citations & References (15)
Invitrogen™
Click-iT™ HPG Alexa Fluor™ 594 Protein Synthesis Assay Kit
The Click-iT™ HPG Alexa Fluor™ 594 Protein Synthesis Assay Kit provides a fast, sensitive, non-toxic, and non-radioactive method for theRead more
| Catalog Number | Quantity |
|---|---|
| C10429 | 25 coverslips or two 96-well plates kit |
Catalog number C10429
Price (MXN)
-
Quantity:
25 coverslips or two 96-well plates kit
The Click-iT™ HPG Alexa Fluor™ 594 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™ 594 azide. The HPG is fed to cultured cells and incorporated into proteins during active protein synthesis. Addition of the Alexa Fluor™ 594 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 complexity
The Click-iT™ HPG Alexa Fluor™ 594 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™ 594 Protein Synthesis Assay Kit and Click-iT™ AHA Alexa Fluor™ 488 HCS Kit, these kits can be used in conjunction for spatial or temporal determination of differences in nascent protein synthesis.
• 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 complexity
The Click-iT™ HPG Alexa Fluor™ 594 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™ 594 Protein Synthesis Assay Kit and Click-iT™ AHA Alexa Fluor™ 488 HCS Kit, these kits 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.
Specifications
ColorRed-Orange
Detection MethodFluorescence
For Use With (Equipment)Fluorescence Microscope, Fluorescent Imager
FormatLiquid
Green FeaturesLess hazardous
Label TypeAlexa Fluor Dyes
Product LineAlexa Fluor, Click-iT
Product TypeProtein Synthesis Assay Kit
Quantity25 coverslips or two 96-well plates kit
Labeling TargetProteins
Label or DyeAlexa Fluor 594
Unit SizeEach
Contents & 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 (15)
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