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MtoZ Biolabs Developed SILAC-based CoIP-MS Service

publication date: Jun 11, 2019
 | 
author/source: MtoZ Biolabs

20190531_1


MtoZ Biolabs, an international contract research organization (CRO) providing advanced proteomics, metabolomics, bioinformatics, and biopharmaceutical analysis services, developed SILAC-based CoIP-MS service to assist researchers study in protein.

Co-Immunoprecipitation (CoIP) is a classical method for studying protein interactions based on the principle that antibodies specifically recognize specific antigens. Immunoprecipitation is often used to test the interaction between proteins under physiological conditions and can be used to identify new interacting proteins by using mass spectrometry (MS).

Mtoz Biolabs uses the SILAC method to label the experimental group and the control group, and then carried out the Co-IP experiment. The immune complex is isolated by the specific reaction between the antigen and the antibody. The LC/MS/MS is used for qualitative/quantitative detection of the proteins in the immune complex. When the content of a certain protein in the experimental group and the control group reached a statistical difference, determining that the protein interacts with the studied protein will greatly reduce the possibility of false positive results of protein interaction. The combination of SILAC technology and Co-IP-MS technology can be used for high-throughput quantitative analysis of protein interaction protein networks in specific situations.

The combination of SILAC introduces quality differences from the source, which reduces experimental system errors, and is rich in quantitative information and high in accuracy. MtoZ Biolabs uses Thermo Fisher's Q ExactiveHF mass spectrometry platform, Orbitrap Fusion mass spectrometry platform, Orbitrap Fusion Lumos mass spectrometry platform combined with Nano-LC to launch a protein interaction analysis service package (SILAC-based CoIP) using SILAC and immuno-precipitation mass spectrometry. -MS). Customers only need to tell them the purpose of their experiment and send out cells, MtoZ Biolabs will be responsible for all subsequent matters of the project, including cell culture, cell labeling, protein extraction, antibody IP, efficiency detection, protease digestion, peptide separation, mass spectrometry, mass spectrometry raw data analysis, bioinformatics analysis, etc.

Characteristics of SILAC-based CoIP-MS

  • preserve protein structure and post-translational modifications
  • Protein interactions are carried out in a natural state to avoid human influence
  • Can separate the natural state of the interacting protein complex
  • High throughput for fast and comprehensive analysis of all interacting proteins
  • Can distinguish non-specific binding proteins compared to the control group, low false positive
  • Ability to compare the relative abundance of each interacting protein

Sample Requirements of SILAC-based CoIP-MS

Protein sample: sample concentration > 0.1 ug/ul; total protein > 300 ug

Animal cells > 1*10^7

Fresh tissue > 100mg

 


About MtoZ Biolabs

MtoZ Biolabs is an international contract research organization (CRO) providing advanced proteomics, metabolomics, bioinformatics, and biopharmaceutical analysis services to researchers in biochemistry, biotechnology, and biopharmaceutical fields. It is specialized in quantitative multiplexed proteomics and metabolomics applications through the establishment of state-of-the-art mass spectrometry platforms, coupled with high-performance liquid chromatography technology.

 

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