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• • How to Use GFP Tags for Co‑immunoprecipitation?

  Protein–protein interactions represent a central component in mapping cellular functional networks. Co-immunoprecipitation (Co-IP), a classical method for interrogating protein interactions, remains widely adopted owing to its procedural simplicity and reliable interpretability. With advances in molecular biology, green fluorescent protein (GFP) tagging has become increasingly prevalent in protein functional studies, and GFP-tag-based Co-IP has emerged as a highly specific and low-background approach ......

• • How to Analyze Protein–Protein Interactions: Key Experimental Approaches

  Protein–protein interactions (PPIs) constitute the foundation of nearly all cellular life processes. They regulate molecular network stability and dynamics, influencing signal transduction, transcriptional regulation, protein degradation, immune responses, and tumorigenesis. In recent years, rapid advances in structural biology, mass spectrometry, and high-throughput screening have transformed PPI research from conventional validation experiments to systematic, network-level, and precision-oriented st......

• • Principles of Mass Spectrometry in Host Cell Protein Identification

  Host cell proteins (HCPs) are impurities derived from production host cells such as CHO, HEK293, or Escherichia coli during the manufacture of recombinant drugs, antibodies, or vaccines. Even at trace levels, these residual proteins may elicit immune responses, compromise drug stability, or pose safety risks. With the continuous elevation of standards in biopharmaceutical development and manufacturing, the accurate identification, quantification, and monitoring of HCPs has become a critical component ......

• • Step-by-Step LC-MS/MS Workflow for Histone Modification Analysis

  Histone post-translational modifications (PTMs) represent a central mechanism underlying chromatin dynamics. These modifications, including acetylation, methylation, phosphorylation, and others, exert profound influence on gene expression, cell fate decisions, and developmental processes. In recent years, liquid chromatography–tandem mass spectrometry (LC-MS/MS) has emerged as a leading approach for characterizing histone modification landscapes owing to its high throughput, sensitivity, and resolutio......

• • How Machine Learning Enhances Data Analysis in Subcellular Proteomics?

  In the post-genomic era, subcellular proteomics has emerged as a key approach for elucidating cellular functions, protein localization, and the mechanisms underlying dynamic regulation. By performing quantitative and qualitative analyses of proteins across distinct subcellular compartments (e.g., nucleus, mitochondria, endoplasmic reticulum), researchers can characterize the spatial dimension of protein function. However, subcellular proteomics data are typically high-dimensional, noisy, and heterogen......

• • How Subcellular Proteomics Drives Biopharmaceutical Development and Precision Medicine?

  Investigating proteins at the cellular level has become central to elucidating biological processes; however, conventional proteomics often overlooks the spatial distribution of proteins within cells. Subcellular proteomics is reshaping this paradigm. By resolving the precise localization and dynamic translocation of proteins within organelles, researchers can construct a cellular functional map that supports drug-target discovery, mechanism-of-action studies, and precision medicine. What Is Subcellu......

• • How to Analyze Signaling Pathways Using Co‑IP-Based Methods?

  In life science research, immune signaling pathways constitute essential biological systems that maintain immune homeostasis and enable the host to respond to pathogenic challenges. These pathways comprise sequential events involving receptor recognition, intracellular signal transmission, and activation of effector molecules, and they participate broadly in processes such as inflammation, antiviral defense, and autoimmunity. Elucidating the dynamic interactions among proteins within immune pathways i......

• • How to Reduce High Background Signals in Co‑IP Assays?

  High background signals frequently occur in Co-immunoprecipitation (Co-IP) experiments and often present as nonspecific bands, substantial interference in Western blotting, or poorly resolved target protein signals. These issues compromise data reliability and may obscure genuine protein–protein interaction events. To improve assay specificity and reproducibility, this article provides a systematic analysis of potential causes and corresponding optimization strategies across four dimensions: experimen......

• • How to Successfully Perform Co-IP for Low-Expression Proteins?

  In life sciences, accurate identification of protein-protein interactions is essential for elucidating cellular signaling pathways, disease mechanisms, and discovering therapeutic targets. Co-immunoprecipitation (Co-IP), a well-established technique for capturing protein interactions, is widely used across biological systems. Nevertheless, Co-IP analysis of low-expression proteins is often hindered by insufficient signals and substantial background interference. Enhancing the enrichment and detection ......

• • How to Combine Co‑IP with Mass Spectrometry for Protein Interaction Analysis?

  Protein–protein interactions (PPIs) constitute the fundamental cellular network that orchestrates essential biological processes including signal transduction, metabolic pathways, and transcriptional regulation. To elucidate the biological role of a protein, it is crucial not only to determine its expression level but also to characterize its interaction partners. Co-immunoprecipitation coupled with mass spectrometry (Co-IP-MS) has emerged as a powerful strategy for mapping endogenous interaction netw......