Quantitative Proteomics Service for Plant Ubiquitination

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Quantitative Proteomics Service for Plant Ubiquitination

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Ubiquitin (Ub) is a highly conserved small molecule protein. The ubiquitin-proteasome system mediates the degradation of most proteins in eukaryotes. In addition, it can also regulate various physiological activities such as cell growth, apoptosis, transcription, immune response, and DNA damage repair.

CD BioSciences utilizes antibodies with a high affinity for lysine residues to enrich ubiquitinated peptide segments in complex samples and combines LC-MS/MS protein quantification methods to achieve a large-scale quantitative analysis of plant ubiquitinated proteins. Our service helps customers understand the mechanisms by which ubiquitinated proteins participate in protein degradation.

Figure 1. Strategies for isolating ubiquitinated proteins/peptides from complex mixtures. – CD BioSciences Figure 1. Strategies for isolating ubiquitinated proteins40/peptides from complex mixtures. (Chen, PC., et al. 2012)

Service Content Description
LC-MS/MS Combined with Ubiquitin Coupling Labeling Service We insert commonly used affinity labels at the N-terminus of ubiquitin molecules and use harsh denaturation conditions (8M urea) to eliminate pollutants. However, this method is not suitable for low-level ubiquitination proteins and is only suitable for qualitative and semi-quantitative experiments.
LC-MS/MS Combined with Ubiquitin Affinity Media Service We enrich ubiquitinated proteins based on the ubiquitin-binding domain (UBD) affinity for ubiquitin chains and can protect the ubiquitin chains from degradation by deubiquitinases. This service is beneficial for the enrichment of some low-level ubiquitinated proteins but has a weak affinity for single ubiquitinated proteins.
LC-MS/MS Combined with Antibody Service We use specific antibodies to enrich ubiquitinated proteins. The purification method of K-ε-GG antibodies can enrich high-level ubiquitination peptide segments, but cannot recognize non lysine ubiquitination modification sites.

Workflow

workflow

Application Fields

We have a professional ubiquitin proteome analysis service platform to help customers solve research problems related to ubiquitination quantification, including

  • The mechanism of plant growth and development.
  • Abnormal regulation of ubiquitin-proteasome degradation pathway.
  • Study on the physiological processes involved in the ubiquitin-proteasome degradation pathway.
  • Study on plant ubiquitination proteomics under light regulation.
  • The effect of hormone treatment on plant ubiquitinated proteins.
  • Plant abiotic stress response mechanism.
  • Plant disease resistance mechanism.
  • Crop improvement.
Notification for Sample Preparation and Handling
Plant leaves, wet weight ≥ 3g/sample.
Plant rhizome, xylem, phloem, etc., wet weight ≥ 5g/sample.
Soft peel ≥ 3g/sample.
Cell samples, cells ≥ 5×107.
Protein extract, concentration > 2mg/mL, total amount ≥ 1mg.
Please use sufficient dry ice for transportation and try to use faster shipping methods to reduce the possibility of sample degradation.

CD BioSciences is a biotechnology company focused on plant protein research, providing a variety of related services and products for environmental and energy solutions. If you are interested in our services, please contact us for more details.

Reference

  1. Chen, PC., et al. (2012). Quantitative proteomics to decipher ubiquitin signaling. Amino Acids. 43:1049–1060.

For research use only, not for clinical use.