What is Protein Microarray?

Protein microarray is an advanced high-throughput technology that allows simultaneous analysis of thousands of proteins’ expression, binding abilities, and functional activities. This technology immobilizes large amounts of proteins onto solid substrates and detects specific interactions and biological activities through labeled probes. Protein microarrays offer high sensitivity, rapid analysis, and automation, making them essential tools in proteomics research and clinical biomarker discovery.

Types of Protein Microarrays

• Analytical Arrays: Use antibodies or aptamers as capture molecules to quantitatively analyze protein expression levels and binding characteristics.
• Reverse Phase Protein Microarrays (RPPA): Spot cell or tissue lysates on chips and detect specific proteins and their post-translational modifications, commonly applied in disease biomarker discovery.
• Functional Protein Microarrays: Immobilize purified proteins to study their interaction activities and enzymatic functions, mimicking physiological protein structures and functions.

Protein Microarray Fabrication Process

1. Substrate Pretreatment and Surface Modification: Selecting suitable slides or membranes and chemically modifying surfaces (e.g., silanization) to enhance protein immobilization efficiency and activity retention.
2. Protein Immobilization: Precisely spotting purified proteins or lysates onto substrates to ensure maintenance of natural conformation and bioactivity.
3. Labeling and Detection: Using fluorescently labeled probes to bind target molecules, followed by laser scanning to quantify protein expression and interactions.

Applications of Protein Microarrays

• Protein Interaction Studies: Revealing complex protein interaction networks including protein-protein, protein-DNA, and protein-small molecule interactions.
• Post-translational Modification Analysis: High-throughput screening of modifications such as phosphorylation and methylation to aid in understanding disease mechanisms.
• Biomarker Identification: Discovering specific protein markers under pathological conditions for early diagnosis and classification of cancer and other diseases.
• Antibody Specificity Analysis: Assessing antibody specificity and affinity to improve diagnostic accuracy.
• Clinical Cancer Protein Expression Profiling: Supporting personalized medicine through targeted protein expression analysis.
• Host-Pathogen Interaction Studies: Elucidating biological mechanisms between bacteria and hosts.

Frequently Asked Questions (FAQ)

What is the difference between protein microarrays and DNA microarrays?

Protein microarrays focus on protein functions and interactions, whereas DNA microarrays mainly analyze gene expression or gene sequences.

What reagents and instruments are needed for protein microarrays?

They primarily include protein purification reagents, surface chemistry treatment agents, fluorescently labeled antibodies or probes, and high-sensitivity laser scanners.

How to choose the appropriate type of protein microarray?

Selection depends on research goals and sample sources, with analytical, reverse phase, or functional arrays chosen for optimal results.

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