User-agent: * Disallow: / Table 1.1. Comparison of various quantitation methodsa | amazing world
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  • Table 1.1. Comparison of various quantitation methodsa


    Protein Lowry Bradford BCA Fluorescence
    α-Casein 0.91
    BSA 1.00 1.00 1.00 1.00
    Calf thymus 1.24 1.10 1.15
    histones
    Chymotryp- 1.52 0.58 0.99
    siongen A
    Cytochrome c 1.39 1.20 1.11
    γ-Globulin 1.07 0.46 0.95
    IgG (human) 1.03
    IgG (mouse) 1.23 0.79
    Lysozyme 1.54 1.00 0.97
    Myoglobin 0.84 1.38 0.92 0.91
    Ovalbumin 0.93 0.52 1.08 0.97
    Ribonuclease A 1.28 0.68
    Soybean trypsin 0.83 0.66
    inhibitor
    Trypsin 1.34 0.34
    Mean ± SD 1.18 ± 0.26 0.81 ± 0.34 1.04 ± 0.10 0.96 ± 0.11
    a Estimated for highly purified proteins; relative to BSA. Data from:
    Peterson GL (1983) Meth Enzymol 91:95; Pierce (1996) Protein assay
    technical protocol; and Invitrogen/Molecular Probe Quant-iT Technical
    Bulletin (2004)
    1.1.1 LOWRY Protein Quantification
    1.1.1.1 Standard Procedure
    This protocol is slightlymodified,with respect to the original paper
    by Lowry et al., to work with smaller volumes. The Folin phenol
    method (Lowry protocol; Table 1.2) is useful in the widest variety
    of experimental applications and is also the least variable with
    different proteins. It is noted that this method, which uses the
    oxidation of aromatic amino acids, is easily disturbed by a lot
    of substances, which are components of the buffer. As a control
    an aliquot of the protein-free buffer in the same volume as the
    protein-containing sample has to be taken as blank1.
    Since the reaction conditionsmay differ fromexperiment to experiment
    and the standard curve is not linear, a couple of standards
    with different amounts of protein between 0 and 100 μg should be
    measured in each analysis. For most purposes a stock solution of
    1 A detailed discussion of Folin–Ciocalteu’s phenol protein determination
    method, especially with respect to possible disturbances and
    troubles and in comparison with the Bradford method, is given by
    Peterson (1996) loc. cit.

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