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Related Concept Videos

Caspases01:24

Caspases

14.2K
Caspase, a family of cysteine proteases, serve as effectors in apoptosis. The ced3 gene in C.elegans was first identified to be involved in apoptosis. This gene encodes the ced-3 caspase that is similar to the interleukin-1-beta converting enzyme or ICE in mammals. In addition to apoptosis, caspases also function in the inflammatory response. Inflammatory caspases are essential in activating pro-inflammatory cytokines that recruit immune cells and block the replication of pathogens inside...
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Quantifying Work02:30

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As a system undergoes a change, its internal energy can change, and energy can be transferred from the system to the surroundings, or from the surroundings to the system.
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Quantifying Heat02:46

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Thermal Energy Microscopically, thermal energy is the kinetic energy associated with the random motion of atoms and molecules. Temperature is a quantitative measure of “hot” or “cold”, which depends on the amount of thermal energy. When the atoms and molecules in an object are moving or vibrating quickly, they have a higher average kinetic energy (KE) (or higher thermal energy), and the object is perceived as “hot”, or it is described as being at a higher temperature. When the...
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SN1 Reaction: Stereochemistry02:15

SN1 Reaction: Stereochemistry

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This lesson provides an in-depth discussion of the stereochemical outcomes in an SN1 reaction.
In the first step of an SN1 reaction, the bond between the electrophilic carbon and the leaving group ionizes to generate the carbocation intermediate. The second step of the mechanism is the nucleophilic attack.
In the formed carbocation, the positively charged carbon is sp2 hybridized with a trigonal planar geometry. As all the three substituents lie on the same plane, a plane of symmetry for the...
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SN1 Reaction: Kinetics02:05

SN1 Reaction: Kinetics

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In an SN2 reaction, the reaction rate depends on both the type of nucleophile and the substrate. A hindered tertiary alkyl halide is practically inert to the SN2 mechanism despite using a strong nucleophile.
However, Sir Christopher Ingold and Edward D. Hughes, who studied the kinetics of various nucleophilic substitution reactions, noticed that a tertiary alkyl halide does undergo a nucleophilic substitution reaction in the presence of a weak nucleophile. While studying the substitution...
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SN1 Reaction: Mechanism02:25

SN1 Reaction: Mechanism

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Kinetic studies of ionization of a tertiary halide in a protic solvent suggest that only the substrate participates in the rate-determining step (slow step). The nucleophile is involved only after the slowest step. The SN1 reaction takes place in a multiple-step mechanism. 
Firstly, the haloalkane ionizes to generate a carbocation intermediate and a halide ion. This heterolytic cleavage is highly endothermic with large activation energy. The ionization of the substrate, facilitated by a...
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Related Experiment Video

Updated: Feb 15, 2026

Detection of Inflammasome Activation and Pyroptotic Cell Death in Murine Bone Marrow-derived Macrophages
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Quantifying Caspase-1 Activity in Murine Macrophages.

Dave Boucher1, Amy Chan1, Connie Ross1

  • 1Institute for Molecular Bioscience and IMB Centre for Inflammation and Disease Research, The University of Queensland, St. Lucia, Australia.

Methods in Molecular Biology (Clifton, N.J.)
|January 12, 2018
PubMed
Summary

This study presents new methods to activate inflammasomes and a direct assay to measure caspase-1 protease activity. This allows for more accurate assessment of inflammatory responses in immune cells.

Keywords:
BiochemistryInflammasomeInflammatory caspasesProtease

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Area of Science:

  • Immunology
  • Molecular Biology
  • Biochemistry

Background:

  • Caspase-1 protease is crucial for inflammasome complexes.
  • Inflammasomes regulate pro-inflammatory cytokines like IL-1β and IL-18.
  • Current caspase-1 activity measurements are indirect and can be unreliable.

Purpose of the Study:

  • To develop methods for activating inflammasomes in murine macrophages.
  • To establish a direct assay for quantifying caspase-1 activity.
  • To provide a more accurate tool for studying inflammatory processes.

Main Methods:

  • Activation of NLRP3, NAIP/NLRC4, and AIM2 inflammasomes in murine macrophages.
  • Development of a simple fluorogenic assay.
  • Direct quantification of cellular caspase-1 activity.

Main Results:

  • Successful elicitation of caspase-1 activity via inflammasome activation.
  • A novel fluorogenic assay was established for direct measurement.
  • The assay provides a straightforward method for quantifying caspase-1 activity.

Conclusions:

  • The described methods enable robust inflammasome activation.
  • The fluorogenic assay offers a direct and simple approach to measure caspase-1 activity.
  • This work provides valuable tools for research into inflammation and related diseases.