Literature DB >> 16146684

Clean Western blot signals from immunoprecipitated samples.

Ashish Lal1, Susan R Haynes, Myriam Gorospe.   

Abstract

We present a strategy that overcomes the high background arising during Western blotting (WB) detection of proteins obtained through immunoprecipitation (IP). Traditional HRP-conjugated secondary antibodies, which detect the denatured heavy and light antibody chains, produce high background that often mask the signals of interest on WBs. Here, we show that HRP-conjugated Protein A and Protein G, which detect almost exclusively intact antibody molecules, can be effectively used to obtain clean and specific WB signals of target proteins.

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Year:  2005        PMID: 16146684      PMCID: PMC1350844          DOI: 10.1016/j.mcp.2005.06.007

Source DB:  PubMed          Journal:  Mol Cell Probes        ISSN: 0890-8508            Impact factor:   2.365


  7 in total

Review 1.  Immunoprecipitation procedures.

Authors:  N E Williams
Journal:  Methods Cell Biol       Date:  2000       Impact factor: 1.441

2.  Concurrent versus individual binding of HuR and AUF1 to common labile target mRNAs.

Authors:  Ashish Lal; Krystyna Mazan-Mamczarz; Tomoko Kawai; Xiaoling Yang; Jennifer L Martindale; Myriam Gorospe
Journal:  EMBO J       Date:  2004-07-15       Impact factor: 11.598

3.  Antiapoptotic function of RNA-binding protein HuR effected through prothymosin alpha.

Authors:  Ashish Lal; Tomoko Kawai; Xiaoling Yang; Krystyna Mazan-Mamczarz; Myriam Gorospe
Journal:  EMBO J       Date:  2005-04-28       Impact factor: 11.598

4.  Protein A from Staphylococcus aureus: the biological significance of its reaction with IgG.

Authors:  A Forsgren; K Nordström
Journal:  Ann N Y Acad Sci       Date:  1974-07-31       Impact factor: 5.691

5.  Immunoprecipitation methods.

Authors:  J J Otto; S W Lee
Journal:  Methods Cell Biol       Date:  1993       Impact factor: 1.441

6.  Detection of protein-protein interactions by coimmunoprecipitation and dimerization.

Authors:  L J Ransone
Journal:  Methods Enzymol       Date:  1995       Impact factor: 1.600

7.  Use of Immunomatrix Methods to Improve Protein-Protein Interaction Detection.

Authors:  M. Walid Qoronfleh; Ling Ren; Daryl Emery; Maria Perr; Barbara Kaboord
Journal:  J Biomed Biotechnol       Date:  2003
  7 in total
  22 in total

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2.  Analysis of the bioactivity of magnetically immunoisolated peroxisomes.

Authors:  Yaohua Wang; Thane H Taylor; Edgar A Arriaga
Journal:  Anal Bioanal Chem       Date:  2011-11-08       Impact factor: 4.142

3.  Rab32 regulates melanosome transport in Xenopus melanophores by protein kinase a recruitment.

Authors:  Minjong Park; Anna S Serpinskaya; Nancy Papalopulu; Vladimir I Gelfand
Journal:  Curr Biol       Date:  2007-11-08       Impact factor: 10.834

4.  Thylakoid FtsH protease contributes to photosystem II and cytochrome b6f remodeling in Chlamydomonas reinhardtii under stress conditions.

Authors:  Alizée Malnoë; Fei Wang; Jacqueline Girard-Bascou; Francis-André Wollman; Catherine de Vitry
Journal:  Plant Cell       Date:  2014-01-21       Impact factor: 11.277

5.  Enzyme-linked enzyme-binding assay for Pin1 WW domain ligands.

Authors:  Ana Y Mercedes-Camacho; Felicia A Etzkorn
Journal:  Anal Biochem       Date:  2010-03-15       Impact factor: 3.365

6.  Binding of Drosophila ORC proteins to anaphase chromosomes requires cessation of mitotic cyclin-dependent kinase activity.

Authors:  Tina Baldinger; Manfred Gossen
Journal:  Mol Cell Biol       Date:  2008-10-27       Impact factor: 4.272

7.  LPS-induced production of TNF-α and IL-6 in mast cells is dependent on p38 but independent of TTP.

Authors:  Thomas Hochdörfer; Christopher Tiedje; Deborah J Stumpo; Perry J Blackshear; Matthias Gaestel; Michael Huber
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8.  Brg1 is required for Cdx2-mediated repression of Oct4 expression in mouse blastocysts.

Authors:  Kai Wang; Satyaki Sengupta; Luca Magnani; Catherine A Wilson; R William Henry; Jason G Knott
Journal:  PLoS One       Date:  2010-05-12       Impact factor: 3.240

Review 9.  An overview of technical considerations for Western blotting applications to physiological research.

Authors:  J J Bass; D J Wilkinson; D Rankin; B E Phillips; N J Szewczyk; K Smith; P J Atherton
Journal:  Scand J Med Sci Sports       Date:  2016-06-05       Impact factor: 4.221

10.  Cooperative folding and ligand-binding properties of LRP6 beta-propeller domains.

Authors:  Chia-Chen Liu; Chelsea Pearson; Guojun Bu
Journal:  J Biol Chem       Date:  2009-03-31       Impact factor: 5.157

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