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Related Concept Videos

Cleavage and Blastulation01:33

Cleavage and Blastulation

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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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Orthogonal Trajectories01:26

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Orthogonal trajectories describe the geometric relationship between two families of curves that intersect each other at right angles. One illustrative case involves a family of parabolas that open sideways along the x-axis. These curves share a common shape but differ by a scaling parameter, resulting in a set of curves that all pass through the origin and widen at different rates.Determining Orthogonal TrajectoriesTo identify the orthogonal trajectories for these parabolas, the first step...
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Isochoric and Isobaric Processes01:21

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A thermodynamic process that occurs at constant volume is called an isochoric process. According to the first law of thermodynamics, heat supplied or removed from the system is partially utilized to perform work and change the internal energy of the system. However, in an isochoric process, the volume remains constant. Hence, the work done by the system is zero. Therefore, the exchange of heat changes the internal energy of the system only. 
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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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Related Experiment Video

Updated: Feb 3, 2026

Automated Sample Multiplexing by using Combined Precursor Isotopic Labeling and Isobaric Tagging cPILOT
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UltraPlex-TMT: Expanding Isobaric Hyperplexing via Orthogonal Protease Cleavage.

Theodoros I Roumeliotis1, Fernando J Sialana1, Jenny Ho2

  • 1The Institute of Cancer Research, Chester Beatty Laboratories, London SW3 6JB, U.K.

Journal of Proteome Research
|February 2, 2026
PubMed
Summary

UltraPlex-TMT enhances quantitative proteomics by doubling sample throughput using TMT/TMTpro labeling and orthogonal digestion. This scalable workflow achieves high protein quantification and reproducibility without custom equipment.

Keywords:
TMThigh-throughput proteomicshyperplexingisobaric labelingorthogonal protease digestionquantitative proteomics

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Area of Science:

  • Proteomics
  • Mass Spectrometry
  • Quantitative Biology

Background:

  • Isobaric labeling techniques like TMT/TMTpro are crucial for multiplexed quantitative proteomics.
  • Current methods face challenges in scaling to higher sample throughput.

Purpose of the Study:

  • To introduce UltraPlex-TMT, a novel workflow for enhanced sample throughput in quantitative proteomics.
  • To demonstrate the scalability and effectiveness of UltraPlex-TMT using hyperplex TMT/TMTpro labeling and orthogonal digestion.

Main Methods:

  • Integration of orthogonal protease digestion (lysine- and arginine-specific) with hyperplex TMT/TMTpro labeling.
  • Benchmarking using a two-species proteome in a pseudo-58-plex design with TMT11plex and TMT18plex.
  • Quantitative analysis using both MS2 and RTS-MS3 acquisition methods.

Main Results:

  • UltraPlex-TMT effectively doubled sample throughput without custom chemistry or instrumentation.
  • Achieved quantification of ~6,000-7,000 proteins per subplex and ~9,000 total proteins with high reproducibility.
  • Demonstrated a trade-off between proteome depth (MS2) and quantification accuracy (MS3), with strong biological concordance between methods.
  • Orthogonal digestion did not introduce systematic quantification bias.

Conclusions:

  • UltraPlex-TMT provides a flexible and scalable foundation for high-throughput proteomics.
  • The workflow supports future advancements towards ultra-high plex scale (200+ samples) and rapid analysis.
  • UltraPlex-TMT facilitates robust proteome profiling with high quantitative accuracy and reproducibility.