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SILAC-based CoIP-MS Service

Co-immunoprecipitation (CoIP) is a classic method to study protein interactions based on the principle that antibodies specifically recognize specific antigens. CoIP is often used to examine the interactions between proteins under physiological conditions and can be used to identify and discover new protein interactions by combining with mass spectrometry (MS).

Our SILAC-based CoIP-MS Service

Creative Proteomics is capable of performing CoIP experiments after nonradioactive isotopic labeling of proteins using the SILAC assay. Then, immune complexes can be isolated and detected utilizing LC-MS/MS based on the specificities between the antigen and the antibody. More specifically, LC-MS/MS statistically detects the differences between proteins of the experimental group and the control group to determine protein interaction and protein networks. This means of detection greatly reduces false positive rates for protein interaction. Hence, the SILAC technology combined with CoIP-MS technology can be used for high-throughput quantitative analysis of protein interaction and networks for various applications.

The combined use of CoIP-MS technology and SILAC assay prevent systematic experimental error and support extensive quantitative information with high accuracy. Creative Proteomics uses Thermo Fisher's Q Exactive HF Mass Spectrometry platform, Orbitrap Fusion Mass Spectrometry platform, and Orbitrap Fusion Lumos Mass Spectrometry platform combined with Nano-LC to launch a SILAC-based CoIP-MS service package for protein interaction analysis. By using this service, customers will only need to send in the sample of investigation, inform objectives of experiments and leave the rest to our team, including but not limited to cell culture, cell labeling, protein extraction, antibody IP, efficiency detection, protease digestion, peptide segment separation, mass spectrometry analysis, mass spectrometry raw data analysis, and bioinformatics analysis.

Workflow of SILAC-based CoIP-MS

Features and Advantages of SILAC-based CoIP-MS

- Protein structure and post-translational modifications are preserved as much as possible
- Protein interactions are carried out in the unmodified confirmation and avoid human factor influences
- Interacting protein complexes can be isolated in unmodified confirmations
- High throughput rapid and comprehensive analysis of all interacting proteins
- Non-specific binding proteins can be identified in comparison with controls at low false positive rates
- Relative abundance of individual interacting proteins can be compared and analyzed

Sample requirements

Sample type (Dry ice transportation)Sample requirements
cell>5×106
Protein extractsTotal protein > 200 μg, concentration >1 μg/μL

Report

1. Experimental procedures
2. Relevant mass spectrometry parameters
3. Details of identified proteins
4. Mass spectrogram
5. Raw data
6. Bioinformatics analysis

* For Research Use Only. Not for use in diagnostic procedures.
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