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Related Experiment Video

Updated: Jun 4, 2026

Microvolume Protein Concentration Determination using the NanoDrop 2000c Spectrophotometer
10:29

Microvolume Protein Concentration Determination using the NanoDrop 2000c Spectrophotometer

Published on: November 4, 2009

Quantifying protein by bicinchoninic Acid.

Richard J Simpson

    CSH Protocols
    |March 2, 2011
    PubMed
    Summary
    This summary is machine-generated.

    This study presents a modified bicinchoninic acid (BCA) assay for protein quantification. The enhanced BCA method offers adaptable procedures for various sample volumes and high-throughput analysis, improving protein detection sensitivity.

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    Area of Science:

    • Biochemistry
    • Analytical Chemistry

    Background:

    • The Lowry assay is a common method for protein quantification.
    • The standard bicinchoninic acid (BCA) assay detects Cu(+) generated from protein-Cu(2+) complexes under alkaline conditions.
    • Traditional BCA assays require large reagent and sample volumes, limiting automation.

    Purpose of the Study:

    • To describe a modified bicinchoninic acid (BCA) assay for protein quantification.
    • To present adaptable procedures for varying sample volumes and automation needs.
    • To enhance the sensitivity and throughput of protein detection.

    Main Methods:

    • The protocol details a variation of the Lowry assay using bicinchoninic acid (BCA).
    • Three procedures are described: Standard, Microprocedure, and 96-Well Microtiter Plate Procedure.

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  • Protein standards are used to generate calibration curves for sample concentration interpolation.

    • Main Results:

    • The modified BCA assay produces a chromophore detectable at 562 nm.
    • The Microprocedure reduces reagent and sample volumes, facilitating automation.
    • The 96-Well Microtiter Plate Procedure allows for high-throughput analysis, potentially reading over 1000 samples per hour with computer interfacing.

    Conclusions:

    • The described BCA assay provides a sensitive and adaptable method for protein quantification.
    • The Microprocedure and 96-Well Microtiter Plate Procedure offer solutions for high-throughput and automated protein analysis.
    • This modified assay enhances the utility of BCA for diverse laboratory applications.