Download - Click-It Technology Illustrated
Click TechnologyApplications in Cell & Molecular Biology
2 12/1/2011 | Life Technologies™ Proprietary and confidential
Technology Basics & Applications
1. Click Technology - Basics & History
2. Click Broad Applications – Cell, in vivo and Molecular Biology applications
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Click Technology – Basics & History
• Click components are bio-orthogonal – very selective & sensitive
• Click components are extremely small and hence bio-friendly
• R1 can be any precursor – Fed metabolically, incorporated enzymatically or chemically
• R2 can be anything such as a dye, bead, enzyme, hapten
• R1 and R2 can be the converse combination as well
Cu+
Extremely slow reaction
Extraordinary temperatures
Rapid reaction – copper catalysis
Room temperature
Patented by Barry Sharpless
Alkyne Azide Stable conjugate
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4 12/1/2011 | Life Technologies™ Proprietary and confidential
Technology Basics & Applications
1. Click Technology - Basics & History
2. Click Broad Applications – Cell, in vivo and Molecular Biology applications
Learn more about Click-It® Technology
5 12/1/2011 | Life Technologies™ Proprietary and confidential
Click Metabolic Precursors – Can Be Fed to Cells & Animals
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• Collect Cells/ Blood/Tissue
• Analyze using Click partner
• Microscopy (Stand alone or HCS)
• Flow cytometry
• Whole well plate readers
• Gel based analysis
Brain Breast Intestine
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Click Metabolic Precursors – Tag & ID Multiple Processes In Cells
Cu+
Monitor nascent DNA synthesis
Monitor new RNA synthesis
Monitor nascent protein synthesis
Monitor post translational modifications
(Alkyne) (Azide)
Cu+
Cu+
Cu+
Cu+
(Stable conjugate)
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Click Metabolic Precursors – Tag & ID Molecules In Wells/Gels
Enzymatic tagging DNA
Enzymatic tagging RNA
Tagging protein in vitro translation
Tagging purified proteins
Cu+
Cu+
Cu+
Cu+
Cu+
(Alkyne) (Azide) (Stable conjugate)
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8 12/1/2011 | Life Technologies™ Proprietary and confidential
Click – Enabling “Cell By Cell” Analysis
• Monitor Cell Proliferation (Click Edu for “S” Phase); Monitor Apoptosis (Click Tunnel)
• Assays are as sensitive as radioactive methods
• Adapted to microscopy, flow and plate base platforms
• Enabling all aspects of biology replication, transcription, translation & PTM
Flow Microscopy Plate readers
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