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A fluorescence-based assay for nanogram quantification of proteins using a protein binding ligand
H Yamini Shrivastava1, Balachandran Unni Nair
1Central Leather Research Institute, Chemical Laboratory, Adyar, 600 020, Chennai, India.
Analytical and Bioanalytical Chemistry
|January 10, 2003
Summary
A novel fluorescent probe enables sensitive protein quantification at nanogram levels. This method overcomes limitations of existing techniques, offering a simple, rapid, and interference-free approach for diverse protein estimations.
Area of Science:
- Biochemistry
- Analytical Chemistry
Background:
- Accurate protein quantification is crucial in biological research.
- Current methods often lack sensitivity or are prone to interference.
Purpose of the Study:
- To develop a sensitive fluorescence-based method for protein estimation at nanogram levels.
- To utilize a Schiff base ligand as a fluorescent probe for protein detection.
Main Methods:
- A Schiff base ligand with donor-acceptor substituents was employed as a fluorescent probe.
- The probe exhibits binding sites for both hydrophobic and hydrophilic protein groups.
- Fluorescence emission changes were monitored in the presence of varying protein concentrations.
Main Results:
- Enhanced fluorescence emission was observed with nanogram levels of protein.
- The method demonstrated a linear response for protein estimation in the range of 66 ng-1 microgram mL(-1).
- The assay showed no interference from buffers, ionic strength, or specific amino acid residues.
Conclusions:
- The developed fluorescence emission-based method is highly sensitive for protein quantification.
- This technique offers a simple, rapid, single-step, and interference-free alternative to existing protein estimation methods.
- The probe's versatility allows its application across different protein classes.