Least-squares A562 curve fit, back-calculated unknowns with dilution factor, extrapolation flags, and detergent/reducing-agent compatibility notes. Educational use only.
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Key facts
Category
Education
Input types
textarea, number
Output type
json
Sample coverage
4
API ready
Yes
Overview
The BCA Assay Calculator fits a least-squares protein standard curve from concentration and A562 readings, then back-calculates unknown protein concentrations using the sample dilution factor. It reports µg/mL and mg/mL results, fit statistics, and extrapolation flags for educational use.
When to use
Quantify unknown protein samples from BCA assay A562 readings.
Fit a standard curve and review its slope, intercept, and R² value.
Correct calculated concentrations for a known sample dilution factor.
How it works
1Enter one protein standard per line as concentration in µg/mL followed by its A562 reading.
2Enter one A562 absorbance value for each unknown sample.
3The calculator fits the standards with least-squares regression and inverts the curve to estimate each unknown.
4It applies the dilution factor and returns concentrations, fit metadata, and flags for readings outside the standard range.
Use cases
Estimate protein concentration in lysate or purified-protein samples from microplate BCA readings.
Compare unknown samples against a standard curve while accounting for pre-assay dilution.
Check curve quality with regression metadata before interpreting calculated concentrations.
Examples
1. Standard-range BCA quantification
Research student
Background
A student measures a BSA standard series from 0 to 2000 µg/mL and records two unknown lysate absorbances.
Problem
They need to convert A562 readings into protein concentrations and confirm that the unknowns fall within the standard curve.
How to use
Enter the standards, add unknown absorbances of 0.735 and 1.640, set the dilution factor to 1, and choose two decimal places.
Enter standards of 0, 200, 500, and 1000 µg/mL with A562 readings of 0.030, 0.230, 0.530, and 1.030. Enter the unknown reading 0.530 and set the dilution factor to 5.
The fitted curve is A562 = 0.001c + 0.03. The assay concentration is 500 µg/mL, and the dilution-corrected sample concentration is 2500 µg/mL, or 2.5 mg/mL.