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Related Concept Videos

Cleavage and Blastulation01:33

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After a large-single-celled zygote is produced via fertilization, the process of cleavage occurs while zygotes travel through the uterine tube. Cleavage is a mitotic cell division that does not result in growth. With each round of successive cell division, daughter cells get increasingly smaller.
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Oxidative Cleavage of Alkenes: Ozonolysis01:46

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In ozonolysis, ozone is used to cleave a carbon–carbon double bond to form aldehydes and ketones, or carboxylic acids, depending on the work-up.
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The reverse of the aldol addition reaction is called the retro-aldol reaction. Here, the carbon–carbon bond in the aldol product is cleaved under acidic or basic conditions to form two molecules of carbonyl compounds. The mechanism of the reaction consists of three steps.
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Alkynes undergo oxidative cleavage in the presence of oxidizing reagents like potassium permanganate and ozone. The triple bond — one σ bond and two π bonds — is completely cleaved, and the alkyne is oxidized to carboxylic acids. When warm and basic aqueous potassium permanganate is used as an oxidizing agent, alkynes are first converted to carboxylate salts via an unstable α-diketone intermediate. Further, a mild acid treatment protonates the carboxylate anions...
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Ethers are generally unreactive and unsuitable for direct nucleophilic substitution reactions since the alkoxy groups are strong bases and, therefore, poor leaving groups. However, ethers readily undergo acidic-cleavage reactions. Ethers can be converted to alkyl halides when heated with strong acids such as HBr and HI in a sequence of two substitution reactions.
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In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
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Related Experiment Video

Updated: Feb 15, 2026

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γ-Secretase Epsilon-cleavage Assay.

Ting-Hai Xu1,2,3, Yan Yan1,2,3, Kaleeckal G Harikumar4

  • 1Key Laboratory of Receptor Research, VARI-SIMM Center, Center for Structure and Function of Drug Targets, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.

Bio-Protocol
|February 3, 2018
PubMed
Summary

This study details a fast and sensitive γ-secretase epsilon-cleavage assay using engineered HEK293 cells. The assay measures γ-secretase activity by quantifying luciferase reporter gene activation following C99 cleavage.

Keywords:
ActivityC99CleavageEpsilon-cleavage assayγ-Secretase

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Background:

  • γ-Secretase is a key enzyme complex involved in intramembrane proteolysis.
  • Dysfunctional γ-secretase activity is implicated in neurodegenerative diseases.
  • Accurate measurement of γ-secretase activity is crucial for drug development.

Purpose of the Study:

  • To present a refined protocol for a γ-secretase epsilon-cleavage assay.
  • To establish a sensitive and rapid method for assessing γ-secretase activity.

Main Methods:

  • Utilized HEK293 cells (HTL cells) with a stably integrated luciferase reporter.
  • Expressed C99 fused to a reversed tetracyclin-inducible activator (rTA).
  • Measured γ-secretase activity via luciferase reporter gene activation upon rTA release.

Main Results:

  • The assay provides a sensitive readout of γ-secretase epsilon-cleavage activity.
  • The protocol is efficient for determining initial C99 cleavage by γ-secretase.

Conclusions:

  • The described cell-based assay is a valuable tool for studying γ-secretase function.
  • This method facilitates research into γ-secretase modulators and related diseases.