Isolation of bacteria-containing phagosomes by magnetic selection.
Lönnbro, Per LU ; Nordenfelt, Pontus LU
and Tapper, Hans
LU
(2008)
In BMC Cell Biology
9(June 27).
- Abstract
- BACKGROUND: There is a growing awareness of the importance of intracellular events in determining the outcome of infectious disease. To improve the understanding of such events, like phagosome maturation, we set out to develop a versatile technique for phagosome isolation that is rapid and widely applicable to different pathogens. RESULTS: We developed two different protocols to isolate phagosomes containing dead or live bacteria modified with small magnetic particles, in conjunction with a synchronized phagocytosis protocol and nitrogen cavitation. For dead bacteria, we performed analysis of the phagosome samples by microscopy and immunoblot, and demonstrated the appearance of maturation markers on isolated phagosomes. CONCLUSION: We have... (More)
- BACKGROUND: There is a growing awareness of the importance of intracellular events in determining the outcome of infectious disease. To improve the understanding of such events, like phagosome maturation, we set out to develop a versatile technique for phagosome isolation that is rapid and widely applicable to different pathogens. RESULTS: We developed two different protocols to isolate phagosomes containing dead or live bacteria modified with small magnetic particles, in conjunction with a synchronized phagocytosis protocol and nitrogen cavitation. For dead bacteria, we performed analysis of the phagosome samples by microscopy and immunoblot, and demonstrated the appearance of maturation markers on isolated phagosomes. CONCLUSION: We have presented detailed protocols for phagosome isolation, which can be adapted for use with different cell types and prey. The versatility and simplicity of the approach allow better control of phagosome isolation, the parameters of which are critical in studies of host-bacteria interaction and phagosome maturation. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1181753
- author
- Lönnbro, Per
LU
; Nordenfelt, Pontus
LU
and Tapper, Hans
LU
- organization
- publishing date
- 2008
- type
- Contribution to journal
- publication status
- published
- subject
- in
- BMC Cell Biology
- volume
- 9
- issue
- June 27
- article number
- 35
- publisher
- BioMed Central (BMC)
- external identifiers
-
- wos:000257835800001
- pmid:18588680
- scopus:47749127370
- ISSN
- 1471-2121
- DOI
- 10.1186/1471-2121-9-35
- language
- English
- LU publication?
- yes
- id
- 01b85614-3a33-432c-a544-912bb45b8b52 (old id 1181753)
- alternative location
- http://www.ncbi.nlm.nih.gov/pubmed/18588680?dopt=Abstract
- date added to LUP
- 2016-04-04 09:25:00
- date last changed
- 2023-04-18 18:54:02
@article{01b85614-3a33-432c-a544-912bb45b8b52,
abstract = {{BACKGROUND: There is a growing awareness of the importance of intracellular events in determining the outcome of infectious disease. To improve the understanding of such events, like phagosome maturation, we set out to develop a versatile technique for phagosome isolation that is rapid and widely applicable to different pathogens. RESULTS: We developed two different protocols to isolate phagosomes containing dead or live bacteria modified with small magnetic particles, in conjunction with a synchronized phagocytosis protocol and nitrogen cavitation. For dead bacteria, we performed analysis of the phagosome samples by microscopy and immunoblot, and demonstrated the appearance of maturation markers on isolated phagosomes. CONCLUSION: We have presented detailed protocols for phagosome isolation, which can be adapted for use with different cell types and prey. The versatility and simplicity of the approach allow better control of phagosome isolation, the parameters of which are critical in studies of host-bacteria interaction and phagosome maturation.}},
author = {{Lönnbro, Per and Nordenfelt, Pontus and Tapper, Hans}},
issn = {{1471-2121}},
language = {{eng}},
number = {{June 27}},
publisher = {{BioMed Central (BMC)}},
series = {{BMC Cell Biology}},
title = {{Isolation of bacteria-containing phagosomes by magnetic selection.}},
url = {{http://dx.doi.org/10.1186/1471-2121-9-35}},
doi = {{10.1186/1471-2121-9-35}},
volume = {{9}},
year = {{2008}},
}