Protein-protein interaction analysis
The function of a protein is functional through multiple protein interactions, so creating an interaction network between proteins can help us understand the composition of a protein complex or the up-and-downstream relationship between different proteins. The most common methods to identify protein interaction is yeast two-hybrid (Y2H) and Affinity-purification mass spectrometry (AP/MS). Y2H uses two proteins to see if they can interact and induce the yeast to express its reporter gene to identify. AP/MS, on the other hand, identifies the composition of the protein complex. AP/MS analysis the whole interactome and can investigate the relationship between proteins, not just to understand if two proteins can interact.
We will elute the immunoprecipitation sample and us in-solution or one-dimension electrophoresis and separate the sample into five regions to do protein identification. After comparing with the database we will know the composition of the proteins in the sample. This method is much more effective than traditional Co-IP strategy.
Service specification
Service specification
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Basic Analysis
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Standard Analysis
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Specification
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1D, 60-90min
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1D, 5 regions
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Note
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Including STRING analysis
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Basic Analysis
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Standard Analysis
Working day not including delivery time: 2 weeks (basic analysis); 3 weeks (standard analysis)
(a) Sample type: Immunoprecipitation product (beads or eluents, please provide buffer constituents for solution sample)
(b) Sample amount: Notspecified (recommended IP starting materials ≥500 ug)
(c) Sample concentration: Not specified.
(d) Platform: Hoefer SE260, 1DLC, LTQ-Orbitrap MS
Please click the link below for the specification of Proteomics service
Sample requirement
1. Summary report (Basic analysis/Standard analysis). pdf
2. Protein-peptide report. xlsx
Q1. If there is no coomassie blue signal after one-dimension electrophoresis, does it mean that there is no protein in the sample and should not be analyzed?
A: No coomassie blue signal does not mean that there is no protein, the amount of protein might be too low for Coomassie blue to detect, so we will still proceed further analyzation.
Q2. Can the samples from immunoprecipitation be eluted before we deliver it?
A: Elution by yourself might elute the antibody and interfere and analyzed results. That is why we strongly suggest doing the elution for you in order to prevent antibody interference.
Q3. What is the difference between in-solution and in-gel for identification?
A: There might still be antibody signal interference in the in-solution sample. Using the in-gel method and separate the sample into five regions can avoid the heavy and light chain of the antibody.
Q4. Do you provide IP service?
A: No, we don't provide IP service. But for protein identification of IP samples, we suggest you let us do elution.
Q5. Will the linker in the protien effect identification results?
A: No, it won't effect.
Q6. If the metod of IP is to label biotin on the protein first then use sreptoavidin to pull down the protein, will biotin and streptoavidin interfere the results?
A: Yes. Biotin and streptoavidin will effect the results. We suggest you provide IP sample and let us do elution. For further information please contact our technician.
Q4. Do you provide IP service?
A: No, we don't provide IP service. But for protein identification of IP samples, we suggest you let us do elution.
Q5. Will the linker in the protien effect identification results?
A: No, it won't effect.
Q6. If the metod of IP is to label biotin on the protein first then use sreptoavidin to pull down the protein, will biotin and streptoavidin interfere the results?
A: Yes. Biotin and streptoavidin will effect the results. We suggest you provide IP sample and let us do elution. For further information please contact our technician.