Purification of caveolae by affinity two-phase partitioning using biotinylated antibodies and NeutrAvidin-dextran
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/138741
- author
- Barinaga-Rementeria Ramírez, Irene LU ; Abedinpour, Parisa LU and Jergil, Bengt LU
- organization
- publishing date
- 2004
- 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
- pmid:15245992
- 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
- 2016-04-01 12:05:51
- date last changed
- 2022-01-26 22:46:18
@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}}, keywords = {{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}}, doi = {{10.1016/j.ab.2004.04.044}}, volume = {{331}}, year = {{2004}}, }