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Related Concept Videos

EDTA: Auxiliary Complexing Reagents01:26

EDTA: Auxiliary Complexing Reagents

EDTA titrations are usually carried out in highly basic conditions, where the fully deprotonated form of EDTA, Y4−, actively complexes with the free metal ions in the solution. Several metal ions precipitate as hydrous oxide (hydroxides, oxides, or oxyhydroxides) under these conditions, lowering the concentration of free metal ions in the solution. For this reason, auxiliary complexing agents or ligands such as ammonia, tartrate, citrate, or triethanolamine are used in EDTA titrations to...
Effects of EDTA on End-Point Detection Methods01:18

Effects of EDTA on End-Point Detection Methods

Different methods, such as visual observance of metal-ion indicators, spectroscopic techniques, and potentiometric methods, can determine the endpoint of an EDTA titration.
In the visual method, metal-ion indicators (metallochromic dyes), which have distinct colors in their free and complex forms, are added to the mixture to signal the titration's end point. They form stable complexes with metal ions, but these complexes are weaker than the corresponding metal–EDTA complexes. As a result, EDTA...
E1 Reaction: Kinetics and Mechanism02:46

E1 Reaction: Kinetics and Mechanism

Here, in contrast to the E2 reaction mechanism, we delve into the aspects of the E1 reaction mechanism, which has two steps: rate-limiting loss of the leaving group and abstraction of the beta hydrogen by a weak base. Typically, the experimental proof for the E1 mechanism is via kinetic studies or isotope studies. While the former demonstrates the first-order kinetics—the dependence of the reaction solely on substrate concentration—the latter proves the abstraction of hydrogen only in the...
E2 Reaction: Kinetics and Mechanism02:45

E2 Reaction: Kinetics and Mechanism

SN2 substitutions and E2 eliminations of alkyl halides proceed via a concerted pathway. While the nucleophile attacks the alpha carbon in SN2 reactions, it functions as a strong base and abstracts a beta hydrogen in the E2 mechanism. The rate-limiting transition state in E2 elimination reactions is characterized by partially broken carbon–hydrogen and carbon–halogen bonds and a partially formed pi bond between the alpha and beta carbons. The beta hydrogen and halide are eliminated...
¹H NMR: Long-Range Coupling01:27

¹H NMR: Long-Range Coupling

The coupling interactions of nuclei across four or more bonds are usually weak, with J values less than 1 Hz. While these are usually not observed in spectra, the presence of multiple bonds along the coupling pathway can result in observable long-range coupling.
In alkenes, spin information is communicated via σ–π overlap, as seen in allylic (four-bond) and homoallylic (five-bond) couplings. These coupling interactions are stronger when the σ bond is parallel to the alkene π orbitals.
NMR Spectroscopy: Spin–Spin Coupling01:08

NMR Spectroscopy: Spin–Spin Coupling

The spin state of an NMR-active nucleus can have a slight effect on its immediate electronic environment. This effect propagates through the intervening bonds and affects the electronic environments of NMR-active nuclei up to three bonds away; occasionally, even farther. This phenomenon is called spin–spin coupling or J-coupling. Coupling interactions are mutual and result in small changes in the absorption frequencies of both nuclei involved. While nuclei of the same element are involved in...

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Updated: Jun 15, 2026

Activated Cross-linked Agarose for the Rapid Development of Affinity Chromatography Resins - Antibody Capture as a Case Study
07:53

Activated Cross-linked Agarose for the Rapid Development of Affinity Chromatography Resins - Antibody Capture as a Case Study

Published on: August 16, 2019

Amine coupling through EDC/NHS: a practical approach.

Marcel J E Fischer1

  • 1Department of Medicinal Chemistry and Chemical Biology, Utrecht Institute for Pharmaceutical Sciences, Utrecht, The Netherlands.

Methods in Molecular Biology (Clifton, N.J.)
|March 11, 2010
PubMed
Summary
This summary is machine-generated.

Optimizing surface plasmon resonance (SPR) immobilization is key for successful interaction analysis. This guide details methods for amine coupling, streptavidin-biotin immobilization, and regeneration to improve experimental outcomes in biomedical research.

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Last Updated: Jun 15, 2026

Activated Cross-linked Agarose for the Rapid Development of Affinity Chromatography Resins - Antibody Capture as a Case Study
07:53

Activated Cross-linked Agarose for the Rapid Development of Affinity Chromatography Resins - Antibody Capture as a Case Study

Published on: August 16, 2019

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The Importance of Correct Protein Concentration for Kinetics and Affinity Determination in Structure-function Analysis

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Engineering Antiviral Agents via Surface Plasmon Resonance
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Engineering Antiviral Agents via Surface Plasmon Resonance

Published on: June 14, 2022

Area of Science:

  • Biomedical research
  • Surface chemistry
  • Biophysical analysis

Background:

  • Surface plasmon resonance (SPR) is a vital technique in biomedical research.
  • Successful SPR analysis hinges on the precise immobilization of interacting molecules onto a sensor surface.
  • Common immobilization failures stem from non-functional or ineffective reactant attachment.

Purpose of the Study:

  • To provide detailed methods for optimizing reactant immobilization in SPR.
  • To discuss common challenges and solutions in surface functionalization for SPR assays.
  • To guide decision-making for enhancing immobilization and regeneration strategies.

Main Methods:

  • Detailed protocols for amine coupling via reactive esters for covalent ligand attachment.
  • Exploration of streptavidin-biotin sandwich immobilization techniques.
  • Strategies for optimizing regeneration conditions to maintain surface activity.

Main Results:

  • Provides a framework for troubleshooting and improving immobilization efficiency.
  • Offers insights into selecting appropriate coupling techniques for diverse molecules (small and large).
  • Enhances understanding of regeneration protocols for assay longevity.

Conclusions:

  • Effective immobilization and regeneration are critical for reliable SPR data.
  • This chapter serves as a practical guide for researchers facing immobilization challenges.
  • Optimized surface chemistry directly impacts the success of SPR-based interaction analysis.