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Protein Quantification with UV-Vis Spectrophotometry

Written by | Published: 09.17.2020

Molecular biology quality control

Protein quantitation is a necessary quality control step in molecular biology workflows to ensure downstream experiments yield accurate results. Microvolume spectrometry enables lab users to sample proteomic materials at very low volumes; allowing labs to preserve precious samples.

Pipette dropping sample onto the Nanodrop One Pedestal for protein quantification

Closeup of pipette dropping sample onto the Nanodrop One Pedestal

Our Thermo Scientific NanoDrop spectrophotometers provide accurate DNA, RNA and protein quantitation with only 1–2 µL of sample. They also enable users to test protein samples as part of batch quality control or as a precursor before additional reactions, quantify the protein content in samples, and identify contaminants that may adversely impact downstream experiments.

Protein quantification e-book

Our recently published Application Compendium eBook for protein quantification and qualification describes how to quantify protein content using either direct or indirect measurements, highlighting the Protein A280 direct application that calculates protein concentration based on the sample absorbance at 280 nm and the protein- and wavelength-specific extinction coefficient. The eBook also details colorimetric assays for indirect measurements and provides protocols for BCA, Pierce Rapid Gold BCA, Bradford, Lowry and the Pierce 660 nm protein assays.

For quantifying peptides that contain Tryptophan and Tyrosine residues in their sequence or peptides that completely lack aromatic amino acids, the eBook steps through the A205 preprogrammed direct absorbance application.

Additionally, the eBook includes:

  •          Information on managing blanking with high absorbing buffers such as RIPA
  •          Details on developing a custom method for hemoglobin measurements
  •          Educational animations
  •          An on-demand webinar

To learn about our spectrophotometers for protein measurements, download your copy the Protein Quantification eBook. Start here.

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