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For workflows utilizing in-solution digestion protocols, it is critical to measure protein concentration following sample lysis using a standard protein assay in order to optimize the ratio of sample/protease (w/w) for digestion. It is also necessary to measure protein/peptide concentration upstream of isobaric labeling to ensure that equal amounts of sample are labeled before mixing. Since most protein assays are not sensitive enough to measure peptide concentration after digestion, the easy-to-use Pierce Colorimetric or Fluorescent peptide assays have been designed specifically to improve the sensitivity and reproducibility of quantitation of peptide mixtures.
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Colorimetric Peptide Assay | Fluorometric Peptide Assay | |
Recommended sample type | Peptide | Peptide |
Measurement | Colorimetric (480 nm) | Fluorescent (Ex. 390, Em. 475 nm) |
Linear range | 15-1000 µg/mL | 5-1000 µg/mL |
Sensitivity | 15 µg/mL | 5 µg/mL |
Minimum sample volume required | 20 µL | 10 µL |
Amount detected at minimum sample volume | 300 ng peptide | 50 ng peptide |
Assay time | 30 min | 5 min |
Key feature | Recommended for TMT-labeled peptides | Recommended for single peptides |
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The BCA Protein Assay become a popular method for colorimetric detection and quantitation of total protein has a unique advantage over other protein assays because of its compatibility with samples that contain up to 5% surfactants (detergents). Compared to most dye-binding methods, the BCA Assay is affected much less by protein compositional differences, providing greater protein-to-protein uniformity.
For Research Use Only. Not for use in diagnostic procedures.