Bradford Protein Content Determination Experiment Report
Objective
The aim of this experiment is to quantify protein concentration via the Bradford assay.
Principle
The Bradford assay is based on the binding of Coomassie Brilliant Blue G-250 to proteins, forming a stable blue complex. The absorbance of this complex peaks at 595 nm when in an acidic environment, enabling the determination of protein concentration.
Methods
1. Standard Curve Development
Prepare BSA solutions with varying concentrations. Add Bradford reagent to each, mix thoroughly, and incubate for 5 minutes. Measure the absorbance at 595 nm.
2. Sample Analysis
Add Bradford reagent to the sample being analyzed, mix thoroughly, and incubate for 5 minutes. Measure absorbance at 595 nm.
Results
Using the absorbance measurements, construct a standard curve that correlates protein concentration with absorbance. Determine the protein concentration of the sample by referencing its absorbance against this standard curve.
The Bradford assay is a rapid, user-friendly, and highly sensitive method for protein quantification, demonstrating specificity and suitability for protein content determination.
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