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Related Concept Videos

Accuracy and Precision01:52

Accuracy and Precision

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Scientists typically make repeated measurements of a quantity to ensure the quality of their findings and to evaluate both the precision and the accuracy of their results. Measurements are said to be precise if they yield very similar results when repeated in the same manner. A measurement is considered accurate if it yields a result that is very close to the true or the accepted value. Precise values agree with each other; accurate values agree with a true value.  Highly accurate...
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Inductively Coupled Plasma-Mass Spectrometry (ICP-MS): Interferences01:20

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Inductively coupled plasma–mass spectrometry (ICP–MS) is a highly selective and sensitive technique for accurate elemental analysis. Though the analysis of ICP–MS mass spectra is comparatively straightforward, it is affected by spectroscopic and non-spectroscopic interferences. Spectroscopic interferences arise when the plasma contains ionic species with an m/z value the same as the analyte ion. Spectroscopic interference can be categorized as isobaric, polyatomic ions, and...
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Inductively Coupled Plasma–Mass Spectrometry (ICP–MS): Overview01:19

Inductively Coupled Plasma–Mass Spectrometry (ICP–MS): Overview

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In inductively coupled plasma–mass spectrometry (ICP–MS), an inductively coupled plasma (ICP) torch is used as an atomizer and ionizer. Solid samples are dissolved and volatilized before being introduced into the high-temperature argon plasma, while solution samples are nebulized and passed through the high-temperature argon plasma. Plasma dissociates the analytes and ionizes their component atoms to form a mixture of positive ions and molecular species. The positive ions are then...
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Uncertainty in Measurement: Accuracy and Precision03:37

Uncertainty in Measurement: Accuracy and Precision

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Scientists typically make repeated measurements of a quantity to ensure the quality of their findings and to evaluate both the precision and the accuracy of their results. Measurements are said to be precise if they yield very similar results when repeated in the same manner. A measurement is considered accurate if it yields a result that is very close to the true or the accepted value. Precise values agree with each other; accurate values agree with a true value. 
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Inductively Coupled Plasma Atomic Emission Spectroscopy: Principle01:19

Inductively Coupled Plasma Atomic Emission Spectroscopy: Principle

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Inductively coupled plasma (ICP) is the most widely used plasma source in atomic emission spectroscopy (AES), also known as Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES). The ICP source, or torch, consists of three concentric quartz tubes with argon gas flowing through them. A spark from a Tesla coil initiates the ionization of argon, generating a high-temperature plasma.
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Immunocytochemistry and Immunohistochemistry01:22

Immunocytochemistry and Immunohistochemistry

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Immunocytochemistry (ICC) and immunohistochemistry (IHC) are techniques that use antibodies to check for specific proteins or antigens in a sample. The technique was first published by Albert Coons in 1941 to detect the presence of pneumococcal antigen in tissue sections from mice infected with Pneumococcus. Immunocytochemistry helps localization of proteins or antigens in individual cells like blood cells, stem cells, etc., while immunohistochemistry does the same for tissue samples.
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Related Experiment Video

Updated: Jan 19, 2026

Isothermal Titration Calorimetry for Measuring Macromolecule-Ligand Affinity
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Isothermal Titration Calorimetry for Measuring Macromolecule-Ligand Affinity

Published on: September 7, 2011

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Obtaining precise and accurate results by ITC.

Lee D Hansen1, Colette Quinn2

  • 1Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT, 84602, USA. lee_hansen@byu.edu.

European Biophysics Journal : EBJ
|September 27, 2019
PubMed
Summary

Achieving accurate isothermal titration calorimetry (ITC) results requires careful experimental design, calibration, and data analysis. Poor practices lead to misleading data, highlighting the need for standardized protocols in protein-ligand binding studies.

Keywords:
AccuracyCalibrationData analysisITCPrecisionStandards

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Collecting Variable-concentration Isothermal Titration Calorimetry Datasets in Order to Determine Binding Mechanisms
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Area of Science:

  • Biophysical Chemistry
  • Biochemistry
  • Analytical Chemistry

Background:

  • Isothermal titration calorimetry (ITC) is a powerful technique for studying molecular interactions, particularly protein-ligand binding.
  • Inaccurate results from ITC studies are frequently reported, stemming from experimental design flaws, instrument calibration issues, and improper data analysis.
  • A recent study claiming to assess ITC's accuracy for protein-ligand binding reactions provided misleading results due to methodological shortcomings.

Purpose of the Study:

  • To identify critical factors influencing the precision and accuracy of isothermal titration calorimetry (ITC) measurements.
  • To analyze common sources of error in ITC experiments, including experimental design, instrument calibration, solution preparation, and data analysis.
  • To propose improvements for obtaining reliable ITC data in the context of protein-ligand binding characterization.

Main Methods:

  • Analysis of a published ITC study on protein-ligand binding to identify experimental and analytical deficiencies.
  • Discussion of essential requirements for robust ITC experimental design, including instrument calibration and parameter optimization.
  • Presentation of experimental results demonstrating achievable precision and accuracy with low-volume ITC instruments.

Main Results:

  • The analyzed study's reported ITC results were misleading due to uncalibrated calorimeters, non-optimized parameters, solution preparation errors, and flawed data analysis.
  • The proposed use of acetazolamide with carbonic anhydrase II as a standard for ITC testing was found to be problematic.
  • The blank correction step was identified as a significant limiting factor for achieving accurate ITC measurements.

Conclusions:

  • Precise and accurate ITC data are contingent upon meticulous experimental design, rigorous instrument calibration, and appropriate data analysis.
  • Standardization of protocols and careful attention to detail, particularly in blank correction, are crucial for reliable ITC studies.
  • Improvements in experimental operations and data handling are necessary to enhance the validity and comparability of ITC results in molecular interaction studies.