Concentration & Prep

Scientific References for Protein Concentration Analysis and Serial Dilution Protocols

The following literature provides the academic and theoretical foundations for determining protein molar extinction coefficients, concentration calculations, and experimental titration protocols. These references can be utilized for research papers, reports, or experimental documentation.

1. Protein Extinction Coefficient Prediction and Concentration Determination

These foundational studies established the algorithms for predicting a protein’s molar absorption coefficient based on its primary amino acid sequence (specifically Tryptophan, Tyrosine, and Cysteine residues).

• Pace, C. N., Vajdos, F., Fee, L., Grimsley, G., & Gray, T. (1995). “How to measure and predict the molar absorption coefficient of a protein.” Protein Science, 4(11), 2411-2423.

  • Application: This is the standard reference for the sequence-based molar extinction coefficient algorithm: ε₂₈₀ = 5500W + 1490Y + 125C.

• Gill, S. C., & von Hippel, P. H. (1989). “Calculation of protein extinction coefficients from amino acid sequence data.” Analytical Biochemistry, 182(2), 319-326.

  • Application: Core theory for protein analysis using amino acid sequence data.

2. Beer-Lambert Law

This fundamental law of physical chemistry defines the linear relationship between the absorbance of a solution and its concentration.

• Beer, A. (1852). “Bestimmung der Absorption des rothen Lichts in farbigen Flüssigkeiten.” Annalen der Physik und Chemie, 162(5), 78-88.

  • Application: The physical origin of concentration determination using the formula A =ε·c·l.

• Swinehart, D. F. (1962). “The Beer-Lambert Law.” Journal of Chemical Education, 39(7), 333.

  • Application: Theoretical background regarding pathlength correction and potential errors in modern analytical chemistry.

3. Protein Molecular Weight and Physical Properties

These references provide the standard values for amino acid molecular weights and the hydration corrections required for sequence-based calculations.

• Cohn, E. J., & Edsall, J. T. (1943). Proteins, Amino Acids and Peptides as Ions and Dipolar Ions. Reinhold Publishing Corp., New York.

  • Application: Fundamental calculations for amino acid residue masses and H2O correction during dehydration synthesis.

• CRC Handbook of Chemistry and Physics. (Latest Edition). Section: Amino Acids and Proteins.

  • Application: Source of standard molecular weight constants for the 20 natural amino acids.

4. Serial Dilution and MST (MicroScale Thermophoresis) Protocols

Theoretical framework for calculating concentration changes in titration experiments and the logic of mixing ligands with target molecules.

• Wienken, C. J., Baaske, P., Rothbauer, U., Braun, D., & Duhr, S. (2010). “Protein binding assay in biological liquids using microscale thermophoresis.” Nature Communications, 1(1), 100.

  • Application: The basis for standard ligand serial dilution protocols and final concentration corrections due to volume changes upon target mixing.

• Segel, I. H. (1976). Biochemical Calculations: How to Solve Mathematical Problems in General Biochemistry. Wiley.

  • Application: Mathematical framework for complex dilution factors and the expanded application of the C1V1 = C2V2 formula.

Citation Guide

When using this software for research purposes, it is recommended to cite it as follows:

In-text citation:

“…the protein concentration and serial dilution table were prepared using ProteinMaker: Concentration & Prep V1.0 (https://proteinmaker.net/wp/concentration-prep/)…”

Full reference:

ProteinMaker (2026). Concentration & Prep V1.0: Advanced Protein Analyzer & Serial Dilution Master. Retrieved from https://proteinmaker.net/wp/concentration-prep/