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Western immunoblotting --- Immunoblotting, Western --- Protein blotting --- Western blot analysis --- Western blotting --- Immunoblotting --- Proteins --- Analysis --- Immunologia --- Proteïnes

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  A key experiment in biomedical research is monitoring the expression of different proteins in order to detect changes that occur in biological systems under different experimental conditions.  The method that is most widely used is the Western blot analysis.  While Western blot is a workhorse in laboratories studying protein expression and has several advantages, it also has a number of significant limitations.  In particular, the method is semi-quantitative with limited dynamic range.  Western blot focuses on a single protein per sample with only a small number of representative samples analyzed in an experiment.  New quantitative tools have been needed for some time to at least supplement, & possibly replace, the Western blot. Mass spectrometric methods have begun to compete with Western blot for routine quantitative analyses of proteins.  One of these methods is based on the tandem mass spectrometry technique of selected reaction monitoring (SRM), which is also called multiple reaction monitoring (MRM).  Selected reaction monitoring is actually an older tandem mass spectrometry technique, first described in the late 70s, that is widely utilized in the quantitative analysis of small molecules like drugs & metabolites.  The use of selected reaction monitoring for the quantitative analysis of proteins has a number of advantages.  Most importantly, it is fundamentally quantitative with a wide dynamic range.  The output of the analysis is a numerical result that can range over several orders of magnitude.  Other advantages include sufficient specificity & sensitivity to detect low abundance proteins in complex mixtures.  Finally, selected reaction monitoring can be multiplexed to allow the quantitative analysis of relatively large numbers of proteins in a single sample in a single experiment.     This Brief will explain both the theoretical & experimental details of the selected reaction monitoring experiment as it is applied to proteins.

Spectrometric and optical chemical analysis --- Genetics --- General biochemistry --- Human biochemistry --- Pathological biochemistry --- Human medicine --- massaspectrometrie --- western blotting --- medische biochemie --- proteomics --- biochemie --- biomedische wetenschappen --- moleculaire biologie --- proteïnen

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Advancements in high-throughput “Omics” techniques have revolutionized plant molecular biology research. Proteomics offers one of the best options for the functional analysis of translated regions of the genome, generating a wealth of detailed information regarding the intrinsic mechanisms of plant stress responses. Various proteomic approaches are being exploited extensively for elucidating master regulator proteins which play key roles in stress perception and signaling, and these approaches largely involve gel-based and gel-free techniques, including both label-based and label-free protein quantification. Furthermore, post-translational modifications, subcellular localization, and protein–protein interactions provide deeper insight into protein molecular function. Their diverse applications contribute to the revelation of new insights into plant molecular responses to various biotic and abiotic stressors.

14-3-3 proteins --- n/a --- targeted two-dimensional electrophoresis --- somatic embryogenesis --- nitrogen metabolism --- subtilase --- Sporisorium scitamineum --- non-orthodox seed --- antioxidant activity --- sweet potato plants infected by SPFMV --- photosynthesis --- B. acuminata petals --- chlorophyll deficiency --- seed proteomics --- imbibition --- pollination --- Sarpo Mira --- qRT-PCR --- holm oak --- tuber phosphoproteome --- isobaric tags for relative and absolute quantitation (iTRAQ) --- Quercus ilex --- nucleotide pyrophosphatase/phosphodiesterase --- lettuce --- ?-subunit --- protein phosphatase --- germination --- drought stress --- pyruvate biosynthesis --- weakening of carbon metabolism --- differential proteins --- heterotrimeric G protein --- organ --- LC-MS-based proteomics --- potato proteomics --- smut --- gel-free/label-free proteomics --- ? subunit --- shotgun proteomics --- 2D --- chloroplast --- proteome functional annotation --- Phalaenopsis --- Clematis terniflora DC. --- wheat --- Dn1-1 --- carbon metabolism --- physiological responses --- Zea mays --- phenylpropanoid biosynthesis --- ISR --- mass spectrometric analysis --- patatin --- leaf --- pea (Pisum sativum L.) --- maize --- ergosterol --- Camellia sinensis --- seed storage proteins --- silver nanoparticles --- elevated CO2 --- metacaspase --- SPV2 and SPVG --- SnRK1 --- MALDI-TOF/TOF --- (phospho)-proteomics --- leaf spot --- rice isogenic line --- wheat leaf rust --- pathway analysis --- phosphoproteome --- sugarcane --- senescence --- Oryza sativa L. --- Arabidopsis thaliana --- heat stress --- gene ontology --- innate immunity --- Pseudomonas syringae --- bolting --- chlorophylls --- shoot --- Simmondsia chinensis --- RT-qPCR --- stresses responses --- Solanum tuberosum --- seeds --- GC-TOF-MS --- sucrose --- proteome --- Puccinia recondita --- cultivar --- Zea mays L. --- secondary metabolism --- ROS --- Ricinus communis L. --- after-ripening --- cadmium --- Stagonospora nodorum --- virus induced gene silencing --- quantitative proteomics --- sweet potato plants non-infected by SPFMV --- affinity chromatography --- population variability --- GS3 --- fungal perception --- ammonium --- transcriptome profiling --- mass spectrometry analysis --- papain-like cysteine protease (PLCP) --- cold stress --- nitrate --- late blight disease --- early and late disease stages --- seed imbibition --- lesion mimic mutant --- protease --- proteome map --- seed dormancy --- petal --- 2-DE proteomics --- 2D DIGE --- root --- Phytophthora infestans --- differentially abundant proteins (DAPs) --- polyphenol oxidase --- degradome --- flavonoid --- 14-3-3 --- caspase-like --- proteomics --- RGG4 --- co-infection --- plasma membrane --- chlorotic mutation --- Medicago sativa --- RGG3 --- glycolysis --- barley --- 2-DE --- protein phosphorylation --- western blotting --- N utilization efficiency --- rice --- plant pathogenesis responses --- high temperature --- data-independent acquisition --- pattern recognition receptors --- vegetative storage proteins --- leaf cell wall proteome --- plant-derived smoke --- iTRAQ --- starch --- proteome profiling --- Morus