Home › Biochemistry › In-vivo Detection of Protein-protein Interactions on Micro-patterned Surfaces
Biochemistry JoVE (Open Access) Citable · DOI

In-vivo Detection of Protein-protein Interactions on Micro-patterned Surfaces

DOI: 10.3791/1969-v
What you'll learn
  • ✓ Design and fabricate micro-patterned surfaces using μ-contact printing
  • ✓ Detect protein-protein interactions in living cells via fluorescence microscopy
  • ✓ Extract quantitative interaction data from high-throughput micropatterned assays
Protocol

Biopharma Insights This video shows experiments with subsequent analysis of protein-protein interactions by the use of micro-patterned surfaces. The approach offers the possibility to detect protein interactions in living cells and combines high throughput capabilities with the possibility to extract quantitative information.

Difficulty
advanced
Total time
~4–6 hours per experiment (including surface preparation, cell incubation, and imaging)
Biosafety
BSL-1

Steps

1
Understand protein interaction detection principles

Review foundational concepts of protein-protein interactions, micropatterning advantages, and high-throughput quantitative screening in living cells.

▶ 00:07
2
Perform μ-contact printing on surfaces

Apply microcontact printing techniques to generate spatially defined protein patterns on experimental substrates for controlled interaction studies.

▶ 02:17
3
Acquire fluorescence microscopy images

Capture high-resolution microscopy images of labeled proteins on micropatterned surfaces to visualize interactions in live or fixed cellular contexts.

▶ 04:15
4
Analyze interaction data quantitatively

Process microscopy data to extract kinetic and affinity parameters for protein-protein interactions from micropatterned arrays.

▶ 05:50
5
Interpret representative results

Examine case study data demonstrating successful detection and quantification of protein interactions on micropatterned surfaces.

▶ 06:27
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