title: Proximity Ligation Assay to Quantify Foxp3 Acetylation in Regulatory T Cells.
abstract: Determining protein acetylation by immunoprecipitation and immunoblotting can be challenging, especially if the tissue of interest is low in quantity, and when good quality acetylation site-specific antibodies are not available. Proximity ligation assays allow a sensitive and quantitative method to assess Foxp3 acetylation in regulatory T cells, with as little as 1.5 × 105cells within two days turnaround time. This method is of potential use in other similar scenarios, when post-translational modifications of a protein of interest need to be determined with only a small amount of sample and in the absence of specific antibodies that can assess the post-translational modification in the protein of interest.
authors.affiliation: Division of Nephrology, Department of Pediatrics, The Children's Hospital of Philadelphia, University of Pennsylvania, 3615 Civic Center Boulevard, 916C Abramson Research Center, Philadelphia, PA, 19104-4318, USA.
authors.forename: Jing
authors.initials: J
authors.lastname: Jiao
authors.affiliation: Laboratory Medicine and Biesecker Center for Pediatric Liver Disease, Division of Transplant Immunology, Department of Pathology, Children's Hospital of Philadelphia, Philadelphia, PA, 19104, USA.
authors.forename: Rongxiang
authors.initials: R
authors.lastname: Han
authors.affiliation: Laboratory Medicine and Biesecker Center for Pediatric Liver Disease, Division of Transplant Immunology, Department of Pathology, Children's Hospital of Philadelphia, Philadelphia, PA, 19104, USA.
authors.forename: Wayne W
authors.initials: WW
authors.lastname: Hancock
authors.affiliation: Division of Nephrology, Department of Pediatrics, The Children's Hospital of Philadelphia, University of Pennsylvania, 3615 Civic Center Boulevard, 916C Abramson Research Center, Philadelphia, PA, 19104-4318, USA. beieru@email.chop.edu.
authors.forename: Ulf H
authors.initials: UH
authors.lastname: Beier
mesh_terms: D000107:Acetylation; D000818:Animals; D000906:Antibodies / Q000737:chemistry; D001681:Biological Assay; D051858:Forkhead Transcription Factors / Q000235:genetics / Q000378:metabolism*; D018189:Immunomagnetic Separation / Q000379:methods*; D047468:Immunoprecipitation; D008198:Lymph Nodes / Q000166:cytology / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D011499:Protein Processing, Post-Translational; D013154:Spleen / Q000166:cytology / Q000378:metabolism; D050378:T-Lymphocytes, Regulatory / Q000166:cytology / Q000378:metabolism*
chemical_list: D000906:Antibodies; D051858:Forkhead Transcription Factors; C498833:Foxp3 protein, mouse
keywords: Immunoblot; Immunopreciptation; Protein acetylation; Proximity ligation assay