title: Optimized semi-quantitative blot analysis in infection assays using the Stain-Free technology.
abstract: Western blots are a commonly used method for protein detection and quantification in biological samples. Compensation of loading variations is achieved by housekeeping protein (HKP) normalization and/or total protein normalization (TPN). However, under infection conditions, HKP normalization, traditionally used in cell biology for quantification of western blots, can be problematic. Binding of microbes to target cells via specific receptors can induce signal transduction events resulting in drastic changes in the level of expression of HKPs. Additionally, samples collected after infection assays will include cellular and microbial proteins altering the analysis with TPN. Here we demonstrate under experimental infection conditions, how a reliable semi-quantitative analysis of proteins in western blots can be achieved using the Stain-Free technology.
authors.affiliation: Max von Pettenkofer-Institut für Hygiene und Medizinische Mikrobiologie, Ludwig-Maximilians-Universität, Pettenkoferstrasse 9a, D-80336 München, Germany.
authors.forename: Anna F
authors.initials: AF
authors.lastname: Zeitler
authors.affiliation: Max von Pettenkofer-Institut für Hygiene und Medizinische Mikrobiologie, Ludwig-Maximilians-Universität, Pettenkoferstrasse 9a, D-80336 München, Germany.
authors.forename: Katrin H
authors.initials: KH
authors.lastname: Gerrer
authors.affiliation: Max von Pettenkofer-Institut für Hygiene und Medizinische Mikrobiologie, Ludwig-Maximilians-Universität, Pettenkoferstrasse 9a, D-80336 München, Germany.
authors.forename: Rainer
authors.initials: R
authors.lastname: Haas
authors.affiliation: Max von Pettenkofer-Institut für Hygiene und Medizinische Mikrobiologie, Ludwig-Maximilians-Universität, Pettenkoferstrasse 9a, D-80336 München, Germany. Electronic address: jimenez@mvp.uni-muenchen.de.
authors.forename: Luisa F
authors.initials: LF
authors.lastname: Jiménez-Soto
mesh_terms: D000942:Antigens, Bacterial / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D001426:Bacterial Proteins / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D015153:Blotting, Western / Q000379:methods*; D002460:Cell Line; D016480:Helicobacter pylori / Q000502:physiology*; D013194:Staining and Labeling
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; C080866:cagA protein, Helicobacter pylori
keywords: Data normalization; Infection biology; Semi-quantitative western blot; Stain-Free technology