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Related Experiment Video

Updated: Jan 22, 2026

Label-Free Immunoprecipitation Mass Spectrometry Workflow for Large-scale Nuclear Interactome Profiling
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Freezing Cell Pellets for Large-Scale Immunoprecipitation.

Larisa Litovchick

    Cold Spring Harbor Protocols
    |July 3, 2019
    PubMed
    Summary

    Freezing large numbers of cells (10^8-10^9) before lysis preserves crucial protein interactions and modifications for immunoprecipitation. This method ensures sample integrity for various scales and time points, simplifying complex experiments.

    Area of Science:

    • Cell biology
    • Biochemistry
    • Molecular biology

    Background:

    • Immunoprecipitation (IP) requires significant cell numbers, posing challenges for large-scale preparation.
    • Protein-protein interactions and post-translational modifications are sensitive to degradation and denaturation during cell lysis.
    • Investigating multiple time points in cell-based studies necessitates robust sample preservation methods.

    Purpose of the Study:

    • To establish a method for large-scale cell preparation prior to immunoprecipitation.
    • To preserve protein integrity and cellular modifications during sample storage.
    • To provide a versatile technique applicable to both adherent and suspension cells across various experimental scales.

    Main Methods:

    • Culturing cells (adherent or suspension) to high viability.

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  • Rinsing cells with phosphate-buffered saline (PBS).
  • Pelleting cells via centrifugation and freezing the resulting pellet at -80°C for long-term storage.

    • Main Results:

    • Successful preservation of protein-protein interactions and post-translational modifications.
    • Demonstrated applicability to large cell numbers (10^8-10^9) and smaller scales.
    • Cell pellets remained viable for storage up to several months at -80°C.

    Conclusions:

    • Cell freezing prior to lysis is an effective strategy for preserving critical molecular components for immunoprecipitation.
    • This method enhances experimental reproducibility and simplifies the workflow for time-course studies.
    • The technique offers a reliable approach for sample preparation in cell biology and biochemical research.