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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3 12/1/2011 | Life Technologies™ Proprietary and confidential

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

Click m

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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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6 12/1/2011 | Life Technologies™ Proprietary and confidential

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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7 12/1/2011 | Life Technologies™ Proprietary and confidential

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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