Stability of reference proteins in human placenta: general protein stains are the benchmark

Autor: Julie Lafond, Dave Lanoix, Cathy Vaillancourt, Andrée-Anne Lacasse, Joey St-Pierre, M. Viau
Přispěvatelé: Institut Armand Frappier (INRS-IAF), Institut National de la Recherche Scientifique [Québec] (INRS)-Réseau International des Instituts Pasteur (RIIP), BioMed Research Centre [Montréal], Institut National de la Recherche Scientifique [Québec] (INRS)-Université du Québec à Trois-Rivières (UQTR)-Université du Québec à Montréal = University of Québec in Montréal (UQAM), Laboratoire de Physiologie materno-foetale, Department of Biological Sciences, Université du Québec à Montréal = University of Québec in Montréal (UQAM), This study was financially supported by a grant from the Natural Sciences and Engineering Research Council of Canada (NSERC # 262011-2009) awarded to C Vaillancourt and from the Canadian Institute of Health Research (CIHR NET 2002-2007) awarded to J Lafond. D Lanoix received tudentships from the Fond de la recherche en Santé du Québec (FRSQ), AA Lacasse from the Fonds québécois de la recherche sur la nature et les technologies (FQRNT) and the NSERC and J St-Pierre from the Réseau Québécois en reproduction (RQR)-NSERC-CREATE and the Fondation Armand Frappier scholarship program.
Jazyk: angličtina
Rok vydání: 2012
Předmět:
Adult
Peptidylprolyl isomerase A
Coefficient of variation
Placenta
Pregnancy Proteins
Young Adult
MESH: Pregnancy
Western blot
Pregnancy
MESH: Protein Stability
medicine
Humans
Diagnostic Techniques
Obstetrical and Gynecological
Glyceraldehyde 3-phosphate dehydrogenase
Serotonin transporter
MESH: Pregnancy Proteins
MESH: Humans
biology
medicine.diagnostic_test
Staining and Labeling
Protein Stability
MESH: Infant
Newborn
Infant
Newborn
Obstetrics and Gynecology
MESH: Adult
Reference Standards
MESH: Placenta
Molecular biology
MESH: Case-Control Studies
Staining
MESH: Diagnostic Techniques
Obstetrical and Gynecological
Pregnancy Complications
MESH: Staining and Labeling
medicine.anatomical_structure
Reproductive Medicine
Biochemistry
MESH: Young Adult
MESH: Pregnancy Complications
Case-Control Studies
[SDV.TOX]Life Sciences [q-bio]/Toxicology
biology.protein
Female
MESH: Reference Standards
Target protein
MESH: Female
Developmental Biology
Zdroj: Placenta
Placenta, Elsevier, 2012, 33 (3), pp.151-6. ⟨10.1016/j.placenta.2011.12.008⟩
ISSN: 0143-4004
DOI: 10.1016/j.placenta.2011.12.008⟩
Popis: International audience; The stability of reference proteins in semi-quantitative Western blot experiments in normal and diseased placenta has never been studied. This study aims to determine the stability of five reference proteins and two general protein stains in placentas from preeclampsia, gestational diabetes mellitus and matched control pregnancies. The stability of the reference proteins was analysed using indicators of inter-group (P value) and intra-group (coefficient of variation) stability. The effect of different normalization strategies was determined by normalizing serotonin transporter (SERT) expression against the different reference protein markers. Results show significant expression variability of β-actin, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), hypoxanthine phosphoribosyltransferase 1 (HPRT1), peptidylprolyl isomerase A (PPIA) and α-tubulin, and that amido black staining is the most stable reference protein marker. Furthermore, results show that SERT expression significantly differs according to the reference protein markers used for its normalization. The present study demonstrated the importance of using stable reference protein markers and normalization strategy in order to get correct results in semi-quantitative Western blot experiments in placental tissues.
Databáze: OpenAIRE
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