Immunoprecipitation of Proteins under Nondenaturing Conditions
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Immunoprecipitation of Proteins under Nondenaturing Conditions. / Lukas, Jiri; Bartek, Jiri; Hansen, Klaus.
Cell Biology, Four-Volume Set. Vol. 4 Elsevier Science Inc., 2006. p. 253-258.Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review
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TY - CHAP
T1 - Immunoprecipitation of Proteins under Nondenaturing Conditions
AU - Lukas, Jiri
AU - Bartek, Jiri
AU - Hansen, Klaus
PY - 2006/12/1
Y1 - 2006/12/1
N2 - Immunoprecipitation of native proteins has proven to be a powerful and widely used approach in addressing questions related to the nature of a single protein or protein complexes existing under different biological conditions. Combined with the recent improvements of protein microsequencing techniques and mass spectrometry, immunoprecipitation also gives the researcher an option to obtain sequence information from unknown proteins identified through coimmnunoprecipitation and thereby collect data on multiprotein complexes. A critical prerequisite for successful analysis of immunoprecipitated native proteins is the quality of the primary antigen-specific antibodies. For the most straightforward interpretation of results, such a reagent should form specific immunocomplexes with the antigen in its native form without dissociating other associated proteins. In several cases it can be advantageous to avoid the scraping of cells into PBS and to perform a more instant lyses procedure based on adding lyses buffer directly to the cell monolayer that has been washed previously three times with ice-cold phosphate-buffered saline (PBS). © 2006
AB - Immunoprecipitation of native proteins has proven to be a powerful and widely used approach in addressing questions related to the nature of a single protein or protein complexes existing under different biological conditions. Combined with the recent improvements of protein microsequencing techniques and mass spectrometry, immunoprecipitation also gives the researcher an option to obtain sequence information from unknown proteins identified through coimmnunoprecipitation and thereby collect data on multiprotein complexes. A critical prerequisite for successful analysis of immunoprecipitated native proteins is the quality of the primary antigen-specific antibodies. For the most straightforward interpretation of results, such a reagent should form specific immunocomplexes with the antigen in its native form without dissociating other associated proteins. In several cases it can be advantageous to avoid the scraping of cells into PBS and to perform a more instant lyses procedure based on adding lyses buffer directly to the cell monolayer that has been washed previously three times with ice-cold phosphate-buffered saline (PBS). © 2006
UR - http://www.scopus.com/inward/record.url?scp=84884878339&partnerID=8YFLogxK
U2 - 10.1016/B978-012164730-8/50216-1
DO - 10.1016/B978-012164730-8/50216-1
M3 - Book chapter
AN - SCOPUS:84884878339
SN - 9780121647308
VL - 4
SP - 253
EP - 258
BT - Cell Biology, Four-Volume Set
PB - Elsevier Science Inc.
ER -
ID: 218255484