Literature DB >> 19588083

Characterization and quality control of antibodies used in ChIP assays.

Géraldine Goens1, Dorina Rusu, Laurent Bultot, Jean-Jacques Goval, Juana Magdalena.   

Abstract

We present here the very robust characterization and quality control (QC) process that we have established for our polyclonal antibodies, which are mainly directed against targets relevant to the epigenetics field such as modified histones, modifying enzymes, and chromatin-interacting proteins. The final purpose of the characterization and QC is to label antibodies as chromatin immunoprecipitation (ChIP) grade. Indeed, the ChIP method is extensively used in epigenetics to study gene regulation and relies on the use of antibodies to select the protein of interest and then precipitate and identify the DNA associated to it. We have optimized in-house all protocols and reagents needed from the first to the last step of antibody characterization. First, following immunizations, the rabbit crude serum is tested for immune response. Whether or not the antibody is specific is determined in further characterizations. Then, only specific antibodies are tested in ChIP using an optimized method which is ideal for antibody screening. Once QC is established for one antibody, it is used to similarly characterize each antibody batch in order to supply researchers in a reproducible manner with validated antibodies. All in all, this demonstrates that we develop epigenetics research tools based on everyday's researcher's needs by providing batch-specific fully characterized ChIP-grade antibodies.

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Year:  2009        PMID: 19588083     DOI: 10.1007/978-1-60327-414-2_2

Source DB:  PubMed          Journal:  Methods Mol Biol        ISSN: 1064-3745


  9 in total

1.  Detailed specificity analysis of antibodies binding to modified histone tails with peptide arrays.

Authors:  Ina Bock; Arunkumar Dhayalan; Srikanth Kudithipudi; Ole Brandt; Philipp Rathert; Albert Jeltsch
Journal:  Epigenetics       Date:  2011-02-01       Impact factor: 4.528

2.  Mapping the regulon of Vibrio cholerae ferric uptake regulator expands its known network of gene regulation.

Authors:  Bryan W Davies; Ryan W Bogard; John J Mekalanos
Journal:  Proc Natl Acad Sci U S A       Date:  2011-07-12       Impact factor: 11.205

3.  Chromatin Immunoprecipitation Followed by Next-Generation Sequencing (ChIP-Seq) Analysis in Ewing Sarcoma.

Authors:  Gwenneg Kerdivel; Valentina Boeva
Journal:  Methods Mol Biol       Date:  2021

Review 4.  Construction and analysis of multiparameter prognostic models for melanoma outcome.

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Journal:  Methods Mol Biol       Date:  2014

5.  Research Resource: Androgen Receptor Activity Is Regulated Through the Mobilization of Cell Surface Receptor Networks.

Authors:  Jordy J Hsiao; Brandon H Ng; Melinda M Smits; Harryl D Martinez; Rohini J Jasavala; Izumi V Hinkson; Damian Fermin; Jimmy K Eng; Alexey I Nesvizhskii; Michael E Wright
Journal:  Mol Endocrinol       Date:  2015-07-16

Review 6.  Single molecule and single cell epigenomics.

Authors:  Byung-Ryool Hyun; John L McElwee; Paul D Soloway
Journal:  Methods       Date:  2014-09-07       Impact factor: 3.608

Review 7.  Multi-layered global gene regulation in mouse embryonic stem cells.

Authors:  Samuel Beck; Bum-Kyu Lee; Jonghwan Kim
Journal:  Cell Mol Life Sci       Date:  2014-09-17       Impact factor: 9.261

Review 8.  Dam it's good! DamID profiling of protein-DNA interactions.

Authors:  Gabriel N Aughey; Tony D Southall
Journal:  Wiley Interdiscip Rev Dev Biol       Date:  2015-09-18       Impact factor: 5.814

Review 9.  A ChIP on the shoulder? Chromatin immunoprecipitation and validation strategies for ChIP antibodies.

Authors:  Fiona C Wardle; Haihan Tan
Journal:  F1000Res       Date:  2015-07-13
  9 in total

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