SILAC-Immunoprecipitation Service

What is SILAC-Immunoprecipitation?

During the process of separating and purifying bait protein-interacting protein complexes, non-specific binding proteins will inevitably appear. These proteins will also be identified by mass spectrometry, becoming false positive interacting proteins, which can affect subsequent selection and verification. The traditional method of judging false positive interacting proteins is to subtract the list of proteins identified by the control group from the list of proteins identified by the bait protein group, and use the remaining proteins as "interacting proteins." However, many true interacting proteins, such as proteins with relatively high abundance, can also appear in the control group, and the traditional method will exclude them.

SILAC-IP immunoprecipitation is a powerful tool for studying protein-protein interactions. It combines two mature technologies: stable isotope labeling of amino acids in cell culture (SILAC) and immunoprecipitation (IP). It can provide high-quality data on protein-protein interactions in a relatively short time.

The SILAC technique first labels the proteins of the experimental group and the control group with different isotopes, i.e., heavy and light chain amino acids. After complex separation, they are mixed in the same tube for enzyme cutting and mass spectrometry. This way, complex proteins can be identified, and the relative content of proteins in the bait protein group and the control group can be determined based on the intensity of the isotope peaks, thereby more accurately identifying interacting proteins.

At Creative Proteomics, we are committed to providing our clients with high-quality and reliable services. We have a proven track record of delivering accurate and reproducible data, and we work closely with our clients to ensure that their specific needs and requirements are met. With our SILAC-IP-MS service, our clients can be confident that they are getting the best possible data to advance their research.

Advantages of SILAC-IP Service

• The ability to study interactions in the native cellular environment. Proteins studied in their natural environment, rather than in isolation, can provide valuable insights into the function of proteins and the pathways in which they are involved.
• Study of weak or transient protein interactions.
• Relatively fast and easy to perform the technique. the SILAC labeling step can be completed in a few days and the IP step in a few hours.

SILAC-IP Technology Platform in Creative Proteomics

Using wild-type cell lines as experimental materials:

• Cultivate cells separately using light and heavy chain amino acids, and extract the labeled proteins in equal amounts.
• Perform Co-IP using normal IgG and specific antibodies against the bait protein for the extracted light chain and heavy chain total proteins separately.
• Mix the Co-IP products of the light and heavy chains.
• Perform trypsin digestion and LC-MS/MS to identify and quantify proteins.

Using overexpressed bait protein cell lines as experimental materials:

• Cultivate normal control cells and overexpressed bait protein cell lines separately using light and heavy chain amino acids (with labels) in equal amounts, and extract the proteins and quantify them again.
• Mix the equal amount of proteins and perform Co-IP using bait protein antibodies.
• Collect the Co-IP protein solution, perform trypsin digestion and LC-MS/MS to identify and quantify proteins.

Using interference bait protein cell lines as experimental materials:

• Cultivate interference bait protein cell lines and normal control cell lines separately using light chain and heavy chain amino acids, and extract the labeled proteins in equal amounts and quantify them again.
• Mix the equal amount of proteins and perform Co-IP using bait protein antibodies.
• Collect the Co-IP protein solution, perform trypsin digestion and LC-MS/MS to identify and quantify proteins.