Colony Immunoblotting Assay for Detection of Bacterial Cell-surface or Extracellular Proteins

Research output: Contribution to journalArticleProfessional

Abstract

This simple protocol describes how to detect antigens from agar-grown bacterial colonies transferred to nitrocellulose using specific antibodies. The protocol is well suitable for detection of bacterial proteins exposed on the cell surface or secreted to the extracellular space and it can be modified also for detection of intracellular proteins. The assay can distinguish bacterial clones with different expression rates (high, medium and low) from colonies that do not express target protein. We used this assay for screening of Mat fimbriae-producing Escherichia coli mutants obtained by mini-Tn5 transposon mutagenesis and immunomagnetic separation (Lehti et al., 2013).
Original languageEnglish
JournalBio-Protocol
Volume3
Issue number17
Publication statusPublished - 5 Sep 2013
MoE publication typeD1 Article in a trade journal

Fields of Science

  • 1183 Plant biology, microbiology, virology

Cite this

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title = "Colony Immunoblotting Assay for Detection of Bacterial Cell-surface or Extracellular Proteins",
abstract = "This simple protocol describes how to detect antigens from agar-grown bacterial colonies transferred to nitrocellulose using specific antibodies. The protocol is well suitable for detection of bacterial proteins exposed on the cell surface or secreted to the extracellular space and it can be modified also for detection of intracellular proteins. The assay can distinguish bacterial clones with different expression rates (high, medium and low) from colonies that do not express target protein. We used this assay for screening of Mat fimbriae-producing Escherichia coli mutants obtained by mini-Tn5 transposon mutagenesis and immunomagnetic separation (Lehti et al., 2013).",
keywords = "1183 Plant biology, microbiology, virology",
author = "Lehti, {Timo Antero} and Benita Westerlund-Wikstr{\"o}m",
year = "2013",
month = "9",
day = "5",
language = "English",
volume = "3",
journal = "Bio-Protocol",
issn = "2331-8325",
publisher = "Bio-protocol LLC",
number = "17",

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Colony Immunoblotting Assay for Detection of Bacterial Cell-surface or Extracellular Proteins. / Lehti, Timo Antero; Westerlund-Wikström, Benita.

In: Bio-Protocol, Vol. 3, No. 17, 05.09.2013.

Research output: Contribution to journalArticleProfessional

TY - JOUR

T1 - Colony Immunoblotting Assay for Detection of Bacterial Cell-surface or Extracellular Proteins

AU - Lehti, Timo Antero

AU - Westerlund-Wikström, Benita

PY - 2013/9/5

Y1 - 2013/9/5

N2 - This simple protocol describes how to detect antigens from agar-grown bacterial colonies transferred to nitrocellulose using specific antibodies. The protocol is well suitable for detection of bacterial proteins exposed on the cell surface or secreted to the extracellular space and it can be modified also for detection of intracellular proteins. The assay can distinguish bacterial clones with different expression rates (high, medium and low) from colonies that do not express target protein. We used this assay for screening of Mat fimbriae-producing Escherichia coli mutants obtained by mini-Tn5 transposon mutagenesis and immunomagnetic separation (Lehti et al., 2013).

AB - This simple protocol describes how to detect antigens from agar-grown bacterial colonies transferred to nitrocellulose using specific antibodies. The protocol is well suitable for detection of bacterial proteins exposed on the cell surface or secreted to the extracellular space and it can be modified also for detection of intracellular proteins. The assay can distinguish bacterial clones with different expression rates (high, medium and low) from colonies that do not express target protein. We used this assay for screening of Mat fimbriae-producing Escherichia coli mutants obtained by mini-Tn5 transposon mutagenesis and immunomagnetic separation (Lehti et al., 2013).

KW - 1183 Plant biology, microbiology, virology

M3 - Article

VL - 3

JO - Bio-Protocol

JF - Bio-Protocol

SN - 2331-8325

IS - 17

ER -