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The importance of adequate fixation for immunofluorescent staining of bovine embryos

Karen Goossens UGent, Leen Vandaele UGent, Eline Wydooghe UGent, M Thys, Jeroen Dewulf UGent, Luc Peelman UGent and Ann Van Soom UGent (2011) REPRODUCTION IN DOMESTIC ANIMALS. 46(6). p.1098-1103
abstract
Immunofluorescent staining is often used to investigate the expression of specific proteins in pre-implantation embryos. The success of this method is determined by the specificity of the antibodies, but also by the protocol used for fixation and permeabilization of the samples. In this study, different fixatives are compared in combination with immunofluorescent staining of caudal-type homeobox 2 (CDX2), fibronectin 1 (FN1) and integrins (ITGs) on bovine blastocysts. For both CDX2 and the ITGs, the outcome of the staining was largely dependent on the fixation methods. Paraformaldehyde fixation was best for the intracellular CDX2 protein, whereas acetone fixation gave the best results for the transmembrane ITGs. No difference was observed for the FN1 staining between samples fixed with paraformaldehyde or acetone. These examples demonstrate that the choice of fixation and permeabilization agents is very important for the outcome of the experiment, and this choice is dictated by the (extra) cellular location of the protein under investigation. Inappropriate fixation and/or permeabilization methods can lead to erroneous conclusions regarding the site and amount of protein expression.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
IN-VITRO, PREIMPLANTATION EMBRYOS, FIBRONECTIN, EXPRESSION
journal title
REPRODUCTION IN DOMESTIC ANIMALS
Reprod. Domest. Anim.
volume
46
issue
6
pages
1098 - 1103
Web of Science type
Article
Web of Science id
000297512400024
JCR category
VETERINARY SCIENCES
JCR impact factor
1.356 (2011)
JCR rank
37/141 (2011)
JCR quartile
2 (2011)
ISSN
0936-6768
DOI
10.1111/j.1439-0531.2011.01770.x
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
2019058
handle
http://hdl.handle.net/1854/LU-2019058
date created
2012-02-03 15:30:32
date last changed
2012-03-08 16:23:05
@article{2019058,
  abstract     = {Immunofluorescent staining is often used to investigate the expression of specific proteins in pre-implantation embryos. The success of this method is determined by the specificity of the antibodies, but also by the protocol used for fixation and permeabilization of the samples. In this study, different fixatives are compared in combination with immunofluorescent staining of caudal-type homeobox 2 (CDX2), fibronectin 1 (FN1) and integrins (ITGs) on bovine blastocysts. For both CDX2 and the ITGs, the outcome of the staining was largely dependent on the fixation methods. Paraformaldehyde fixation was best for the intracellular CDX2 protein, whereas acetone fixation gave the best results for the transmembrane ITGs. No difference was observed for the FN1 staining between samples fixed with paraformaldehyde or acetone. These examples demonstrate that the choice of fixation and permeabilization agents is very important for the outcome of the experiment, and this choice is dictated by the (extra) cellular location of the protein under investigation. Inappropriate fixation and/or permeabilization methods can lead to erroneous conclusions regarding the site and amount of protein expression.},
  author       = {Goossens, Karen and Vandaele, Leen and Wydooghe, Eline and Thys, M and Dewulf, Jeroen and Peelman, Luc and Van Soom, Ann},
  issn         = {0936-6768},
  journal      = {REPRODUCTION IN DOMESTIC ANIMALS},
  keyword      = {IN-VITRO,PREIMPLANTATION EMBRYOS,FIBRONECTIN,EXPRESSION},
  language     = {eng},
  number       = {6},
  pages        = {1098--1103},
  title        = {The importance of adequate fixation for immunofluorescent staining of bovine embryos},
  url          = {http://dx.doi.org/10.1111/j.1439-0531.2011.01770.x},
  volume       = {46},
  year         = {2011},
}

Chicago
Goossens, Karen, Leen Vandaele, Eline Wydooghe, M Thys, Jeroen Dewulf, Luc Peelman, and Ann Van Soom. 2011. “The Importance of Adequate Fixation for Immunofluorescent Staining of Bovine Embryos.” Reproduction in Domestic Animals 46 (6): 1098–1103.
APA
Goossens, Karen, Vandaele, L., Wydooghe, E., Thys, M., Dewulf, J., Peelman, L., & Van Soom, A. (2011). The importance of adequate fixation for immunofluorescent staining of bovine embryos. REPRODUCTION IN DOMESTIC ANIMALS, 46(6), 1098–1103.
Vancouver
1.
Goossens K, Vandaele L, Wydooghe E, Thys M, Dewulf J, Peelman L, et al. The importance of adequate fixation for immunofluorescent staining of bovine embryos. REPRODUCTION IN DOMESTIC ANIMALS. 2011;46(6):1098–103.
MLA
Goossens, Karen, Leen Vandaele, Eline Wydooghe, et al. “The Importance of Adequate Fixation for Immunofluorescent Staining of Bovine Embryos.” REPRODUCTION IN DOMESTIC ANIMALS 46.6 (2011): 1098–1103. Print.