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Related Experiment Videos

A quantitative Western Blot method for protein measurement.

C A Dennis-Sykes, W J Miller, W J McAleer

    Journal of Biological Standardization
    |October 1, 1985
    PubMed
    Summary
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    This study introduces a sensitive radioimmunologic assay for quantifying low protein levels (20-150 ng) in recombinant vaccines. The method accurately measures recombinant protein percentages in complex mixtures like bacterial and yeast cell lysates.

    Area of Science:

    • Biochemistry
    • Immunology
    • Biotechnology

    Background:

    • Accurate quantification of proteins is crucial for recombinant vaccine development.
    • Existing methods may lack sensitivity for low-abundance proteins.

    Purpose of the Study:

    • To evaluate a radioimmunologic assay for quantifying small protein amounts (20-150 ng) in recombinant vaccines.
    • To assess the utility of this method for analyzing cell lysates.

    Main Methods:

    • Utilized SDS-PAGE and Western Blot for protein separation and transfer.
    • Employed specific antibodies and radiolabeled Protein A for detection.
    • Quantified protein using a gamma counter after excising from the blot.

    Main Results:

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  • The assay demonstrated sensitivity for detecting 20-150 ng of protein.
  • Autoradiograms served as accurate templates for protein localization.
  • The technique proved effective for evaluating recombinant protein percentage in cell lysates.

    • Conclusions:

    • The radioimmunologic assay is a valuable tool for precise protein quantification in recombinant vaccine research.
    • This method is particularly useful for assessing the purity of recombinant proteins in complex biological mixtures.