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Updated: Oct 19, 2025

Linearization of the Bradford Protein Assay
Published on: April 12, 2010
Measurement Assurance Program Transmittance Standards for Spectrophotometric Linearity Testing: Preparation and
Kenneth L Eckerle1, Victor R Weidner1, Jack J Hsia1
1National Bureau of Standards, Washington, DC 20234.
Abstract:
A Measurement Assurance Program for spectrophotometry is being established in order to assist laboratories involved in spectrophotometric calibrations. This paper deals with the preparation and calibration of neutral density glass filters for checking the linearity of photometric response, as applied to spectral transmittance measurements. Several sets of filters were prepared from suitable neutral glass to provide nominal transmittances of 92, 70, 50, 25, 10, 1, and 0.1% at a wavelength of 548.5 nm. These filter sets will be available in three sizes: these are, 38 mm diameter aperture in 51 × 51 mm holder, 25 mm diameter aperture in 51 × 28 mm holder, and 30 × 8 mm aperture in a cuvette holder. The filters were calibrated for spectral transmittance on the NBS Reference Spectrophotometer for high accuracy transmittance measurements. Measurements were made with a 1.5 nm passband collimated sample beam. The filters were checked for uniformity and measurements were made to determine the effects of sample beam polarization. The transmittance data for the wavelength range of interest were analyzed by statistical methods to determine the effects of passband for a range of 1.5 nm to 10.5 nm passband. The results of these measurements are presented in tabular and graphical detail for the master filter set.
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