Improved immunomatrix methods to detect protein:protein interactions

Ren, Ling; Emery, Daryl J.; Kaboord, Barbara; Chang, Edith; Qoronfleh, M. Walid

doi:10.1016/s0165-022x(03)00105-2

Cited by 25 publications

(13 citation statements)

References 23 publications

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“…Thus, the recovered IgGs were subsequently used for the purification and characterization of parasite antigens by immunoaffinity. The traditional method of immunoprecipitation by incubating an antibody with a protein sample and subsequently binding it to a protein A or G agarose matrix serves to efficiently recover target antigens Ren et al 2003;Roque et al 2007). However, immunoprecipitation followed by weak binding to agarose through protein A or G has certain intrinsic shortcomings such as the loss of insoluble Fig.…”

Section: Discussionmentioning

confidence: 99%

Plasmodium yoelii blood-stage antigens newly identified by immunoaffinity using purified IgG antibodies from malaria-resistant mice

et al. 2012

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Section: Discussionmentioning

confidence: 99%

Plasmodium yoelii blood-stage antigens newly identified by immunoaffinity using purified IgG antibodies from malaria-resistant mice

et al. 2012

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“…Immunoprecipitation approaches, used alone or in association with other separation methods, was extensively described as probes for elucidating some aspects of signal transduction [155] as well as for the analysis of phosphotyrosyl proteins in cerebrospinal fluid [156] and the mapping of phosphorylation sites from human T-cells [157]. The immunoprecipitation principle is also interesting for investigating the formation of protein-protein complexes [158][159][160], and therefore contributing to the elucidation of some pathways.…”

Section: Immunoprecipitationmentioning

confidence: 99%

Prefractionation techniques in proteome analysis: The mining tools of the third millennium

et al. 2005

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The present review deals with prefractionation protocols used in proteomic investigation in preparation for mass spectrometry (MS) or two-dimensional electrophoresis (2-DE) map analysis. Briefly, reported methods focus on cell organelle differential centrifugation and on chromatographic approaches, to continue in extenso with a panoply of electrophoretic methods. In the case of chromatography, procedures useful as a prefractionation step, including affinity, ion-exchange, and reversed-phase resins, revealed several hundreds of new species, previously undetected in unfractionated samples. Novel chromatographic prefractionation methods are also discussed such as a multistaged fractionation column, consisting in a set of immobilized chemistries, serially connected in a stack format (an assembly of seven blocks), each capable of harvesting a given protein population. Such a method significantly simplifies the complexity of treated samples while concentrating species, all resulting in a larger number of visible proteins by MS or 2-DE. Electrophoretic prefractionation protocols include all those electrokinetic methodologies which are performed in free solution, essentially all relying on isoelectric focusing steps (although some approaches based on gels and granulated media are also discussed). Devices associated with electrophoretic separation are multichamber apparatus, such as the multicompartment electrolyzers equipped with either isoelectric membranes or with isoelectric beads. Multicup device electrophoresis and several others, exploiting the conventional technique of carrier ampholyte focusing, are reviewed. This review also reports approaches for sample treatments in order to detect low-abundance species. Among others, a special emphasis is made on the reduction of concentration difference between proteins constituting a sample. This latter consists in a library of combinatorial ligands coupled to small beads. Such a library comprises hexameric ligands composed of 20 amino acids, resulting in millions of different structures. When these beads are impregnated with complex proteomes (e.g., human sera) of widely differing protein compositions, they are able to significantly reduce the concentration differences, thus greatly enhancing the possibility to evidence low-abundance species. It is felt that this panoply of methods could offer a strong step forward in "mining below the tip of the iceberg" for detecting the "unseen proteome".

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“…Using co-immunoprecipitation, we have now identified MGAT2 as another DGAT2-interacting protein. Co-immunoprecipitation is a classic technique that has been routinely used to study protein-protein interactions (41,42). Using this approach, we found that DGAT2 co-immunoprecipitated with MGAT2 both in HEK-293T and McArdle RH7777 cells.…”

Section: Discussionmentioning

confidence: 90%

Diacylglycerol Acyltransferase-2 (DGAT2) and Monoacylglycerol Acyltransferase-2 (MGAT2) Interact to Promote Triacylglycerol Synthesis

Jin

McFie

Banman

et al. 2014

Journal of Biological Chemistry

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Background: DGAT2 catalyzes the final and committed step of triacylglycerol (TG) biosynthesis. Results: Co-immunoprecipitation experiments and in situ proximity ligation assays showed that DGAT2 and MGAT2 interact. Conclusion: DGAT2 can utilize diacylglycerol generated by MGAT2 for TG synthesis. Significance: These findings provide insight into the spatial arrangement of enzymes involved in triacylglycerol biosynthesis.

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Improved immunomatrix methods to detect protein:protein interactions

Cited by 25 publications

References 23 publications

Plasmodium yoelii blood-stage antigens newly identified by immunoaffinity using purified IgG antibodies from malaria-resistant mice

Plasmodium yoelii blood-stage antigens newly identified by immunoaffinity using purified IgG antibodies from malaria-resistant mice

Prefractionation techniques in proteome analysis: The mining tools of the third millennium

Diacylglycerol Acyltransferase-2 (DGAT2) and Monoacylglycerol Acyltransferase-2 (MGAT2) Interact to Promote Triacylglycerol Synthesis

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