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Purification of caveolae by affinity two-phase partitioning using biotinylated antibodies and NeutrAvidin-dextran

Barinaga-Rementeria Ramírez, Irene LU ; Abedinpour, Parisa LU and Jergil, Bengt LU (2004) In Analytical Biochemistry 331(1). p.17-26
Abstract
A new concept for affinity two-phase partitioning was tested. The partitioning was based on the interaction of target membranes with a primary antibody which, in turn, interacted with a biotinylated secondary antibody and NeutrAvidin-dextran in a poly(ethylene glycol)/dextran two-phase system. Caveolae selectively redistributed from the top phase to the NeutrAvidin-dextran-containing bottom phase by employing anti-caveolin as the primary antibody. This immunoaffinity approach was more selective than the established sucrose gradient centrifugation method and resulted in highly purified caveolae from Triton X-100-treated liver and lung plasma membranes. The same approach, employing other selective primary antibodies, should facilitate the... (More)
A new concept for affinity two-phase partitioning was tested. The partitioning was based on the interaction of target membranes with a primary antibody which, in turn, interacted with a biotinylated secondary antibody and NeutrAvidin-dextran in a poly(ethylene glycol)/dextran two-phase system. Caveolae selectively redistributed from the top phase to the NeutrAvidin-dextran-containing bottom phase by employing anti-caveolin as the primary antibody. This immunoaffinity approach was more selective than the established sucrose gradient centrifugation method and resulted in highly purified caveolae from Triton X-100-treated liver and lung plasma membranes. The same approach, employing other selective primary antibodies, should facilitate the purification also of other membrane fractions. (C) 2004 Elsevier Inc. All rights reserved. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
EM, Isolation, 5′-Nucleotidase, Caveolin, Immunoaffinity partitioning, Avidin, Biotin, Two-phase system, Membrane
in
Analytical Biochemistry
volume
331
issue
1
pages
17 - 26
publisher
Elsevier
external identifiers
  • pmid:15245992
  • wos:000222697200002
  • scopus:3042756160
ISSN
1096-0309
DOI
10.1016/j.ab.2004.04.044
language
English
LU publication?
yes
id
c484b128-4964-4f5f-a3f4-8137bb720131 (old id 138741)
date added to LUP
2007-07-06 14:23:03
date last changed
2017-01-01 04:50:00
@article{c484b128-4964-4f5f-a3f4-8137bb720131,
  abstract     = {A new concept for affinity two-phase partitioning was tested. The partitioning was based on the interaction of target membranes with a primary antibody which, in turn, interacted with a biotinylated secondary antibody and NeutrAvidin-dextran in a poly(ethylene glycol)/dextran two-phase system. Caveolae selectively redistributed from the top phase to the NeutrAvidin-dextran-containing bottom phase by employing anti-caveolin as the primary antibody. This immunoaffinity approach was more selective than the established sucrose gradient centrifugation method and resulted in highly purified caveolae from Triton X-100-treated liver and lung plasma membranes. The same approach, employing other selective primary antibodies, should facilitate the purification also of other membrane fractions. (C) 2004 Elsevier Inc. All rights reserved.},
  author       = {Barinaga-Rementeria Ramírez, Irene and Abedinpour, Parisa and Jergil, Bengt},
  issn         = {1096-0309},
  keyword      = {EM,Isolation,5′-Nucleotidase,Caveolin,Immunoaffinity partitioning,Avidin,Biotin,Two-phase system,Membrane},
  language     = {eng},
  number       = {1},
  pages        = {17--26},
  publisher    = {Elsevier},
  series       = {Analytical Biochemistry},
  title        = {Purification of caveolae by affinity two-phase partitioning using biotinylated antibodies and NeutrAvidin-dextran},
  url          = {http://dx.doi.org/10.1016/j.ab.2004.04.044},
  volume       = {331},
  year         = {2004},
}