Accurate measurement of protein concentrations in mixtures is challenging because it is problematic to know the extinction coefficient of a mixture and because proteins respond to protein assays (such as the Bradford assay) differently. Snake venoms are variable mixtures of different proteins. Because of the variability, the use of extinction coefficients would not be possible even if the extinction coefficients of all the specific proteins were identified. Here we show that North American, African and Asian snake venoms all demonstrate approximately the same response to the linearized Bradford assay on a dry weight basis, even though these venoms have different protein compositions. The response of �-lactaglobulin to the linearized Bradford assay is approximately the same as the snake venoms and different from that of other common proteins standards such as bovine serum albumin (BSA) or Ovalbumin (OVA). The concentration of �-lactaglobulin can be determined spectraphotometrically. This allows for the accurate and reproducible preparation of standard curves. The combination of the linearized Bradford and �-lactaglobulin standards suggests the possibility of a reproducible and universal assay for snake venoms around the world, further assisting individuals creating antivenoms.

Accurate measurement of protein concentrations in snake venoms using �-lactaglobulin as a standard protein in the linearized Bradford assay

247th National Meeting (2014)

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