Literature DB >> 8024796

Immunoaffinity purification of FLAG epitope-tagged bacterial alkaline phosphatase using a novel monoclonal antibody and peptide elution.

B L Brizzard1, R G Chubet, D L Vizard.   

Abstract

The FLAG epitope is an eight amino acid peptide (AspTyrLysAspAspAspAspLys) that is useful for immunoaffinity purification of fusion proteins. A monoclonal antibody (anti-FLAG M1) that binds the FLAG epitope in a calcium-dependent manner and requires an N-terminal FLAG sequence has been described previously. We describe the use of a second anti-FLAG monoclonal antibody (anti-FLAG M2) in immunoaffinity purification of N-terminal Met-FLAG and C-terminal FLAG fusion to bacterial alkaline phosphatase. Although binding of an anti-FLAG M2 monoclonal antibody to the FLAG epitope is not calcium-dependent, bound fusion proteins can be eluted by competition with FLAG peptide.

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Year:  1994        PMID： 8024796

Source DB:  PubMed          Journal:  Biotechniques        ISSN： 0736-6205            Impact factor:   1.993


  55 in total

1.  Role and mechanism of action of C. PvuII, a regulatory protein conserved among restriction-modification systems.

Authors:  R M Vijesurier; L Carlock; R M Blumenthal; J C Dunbar
Journal:  J Bacteriol       Date:  2000-01       Impact factor: 3.490

2.  Multiple tandem epitope tagging for enhanced detection of protein expressed in mammalian cells.

Authors:  L Zhang; R Hernan; B Brizzard
Journal:  Mol Biotechnol       Date:  2001-11       Impact factor: 2.695

3.  Type IV-like pili formed by the type II secreton: specificity, composition, bundling, polar localization, and surface presentation of peptides.

Authors:  Guillaume Vignon; Rolf Köhler; Eric Larquet; Stéphanie Giroux; Marie-Christine Prévost; Pascal Roux; Anthony P Pugsley
Journal:  J Bacteriol       Date:  2003-06       Impact factor: 3.490

4.  Roles for receptors, pheromones, G proteins, and mating type genes during sexual reproduction in Neurospora crassa.

Authors:  Hyojeong Kim; Sara J Wright; Gyungsoon Park; Shouqiang Ouyang; Svetlana Krystofova; Katherine A Borkovich
Journal:  Genetics       Date:  2012-01-31       Impact factor: 4.562

Review 5.  Tools used to study how protein complexes are assembled in signaling cascades.

Authors:  Susan Dwane; Patrick A Kiely
Journal:  Bioeng Bugs       Date:  2011-09-01

6.  The ubiquitin-like protein Smt3p is activated for conjugation to other proteins by an Aos1p/Uba2p heterodimer.

Authors:  E S Johnson; I Schwienhorst; R J Dohmen; G Blobel
Journal:  EMBO J       Date:  1997-09-15       Impact factor: 11.598

7.  Infectivity and neutralization of simian immunodeficiency virus with FLAG epitope insertion in gp120 variable loops.

Authors:  Melissa E Laird; Ronald C Desrosiers
Journal:  J Virol       Date:  2007-08-08       Impact factor: 5.103

8.  Loop 1 dynamics in smooth muscle myosin: isoform specific differences modulate ADP release.

Authors:  Justin A Decarreau; Lynn R Chrin; Christopher L Berger
Journal:  J Muscle Res Cell Motil       Date:  2011-06-05       Impact factor: 2.698

9.  The molecular basis for relative physiological functionality of the ADP/ATP carrier isoforms in Saccharomyces cerevisiae.

Authors:  Christopher P Smith; Peter E Thorsness
Journal:  Genetics       Date:  2008-06-18       Impact factor: 4.562

Review 10.  Strategies for achieving high-level expression of genes in Escherichia coli.

Authors:  S C Makrides
Journal:  Microbiol Rev       Date:  1996-09
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