Following extraction, peptide concentration of each sample was measured in duplicate with the Pierce Quantitative Fluorescent Peptide Assay (Thermo Scientific, Rockford, IL, USA). Peptides were labeled by an amine-reactive fluorescent detection reagent, and the fluorescence intensity was detected at Ex390nm/Em475nm with a microplate reader (SpectraMax M5, Molecular Devices, San Jose, CA, USA).
Pierce quantitative fluorescent peptide assay
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Pierce Quantitative Fluorescent Peptide Assay is a fluorescence-based method for the quantitative analysis of peptides. It provides a sensitive and accurate way to measure peptide concentrations in samples.
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2 protocols using pierce quantitative fluorescent peptide assay
1
Fluorescent Peptide Quantification
2
Protein Extraction and Quantification
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The samples were added with Lysis buffer (EasyPep™ Mini MS Sample Prep Kit, Thermo Scientific, USA) to extract the proteins. The lysates were then centrifuged at 16,000 × g for 10 min at 4 oC. The supernatant of the respective samples was aliquot into new tubes. Protein quantitation was done using DC Protein Assay (Bio-Rad Laboratories Inc., USA) for control sample. The protein solutions were then reduced, alkylated, digested and cleaned-up according to the manufacturer’s instruction (EasyPep™ Mini MS Sample Prep Kit, Thermo Scientific, USA).
Peptides were reconstituted in 2% Acetonitrile, 0.1% formic acid in water and peptide concentration was determined by Thermo Scientific™ Pierce™ Quantitative Fluorescent Peptide Assay.
Peptides were reconstituted in 2% Acetonitrile, 0.1% formic acid in water and peptide concentration was determined by Thermo Scientific™ Pierce™ Quantitative Fluorescent Peptide Assay.
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