Cold Spring Harbor protocols最新文献_第8页

Amino Acid Quantification from Maize Tissues. 玉米组织氨基酸定量分析。

Cold Spring Harbor protocols Pub Date : 2024-12-23 DOI: 10.1101/pdb.top108440

Huda Ansaf, Abou Yobi, Ruthie Angelovici

{"title":"Amino Acid Quantification from Maize Tissues.","authors":"Huda Ansaf, Abou Yobi, Ruthie Angelovici","doi":"10.1101/pdb.top108440","DOIUrl":"https://doi.org/10.1101/pdb.top108440","url":null,"abstract":"Amino acid analysis is a vital part of analytical biochemistry. The increasing demand for low nitrogen fertilization and for plant-based diets with balanced amino acid levels and composition have made it crucial to develop reliable, fast, and affordable methods for analyzing amino acids in plants. As maize accounts for 43% of global cereal production, improving the amino acid composition of its kernels (i.e., seeds) is critically important for meeting the dietary requirements of humans and livestock. Moreover, amino acid quantification in maize leaves is necessary for improving yield prediction, stress sensing, and nitrogen use efficiency. Many amino acid quantification methods use reverse-phase high-pressure liquid chromatography and gas chromatography approaches to assess the amino acid content of maize tissues. Historically, these techniques involved the use of chemical derivatization, a chemical reaction that alters the properties of a compound to make it detectable or more sensitive to detection. Although accurate, these methods are time-consuming, expensive, and unsuitable for large populations. Here, we introduce two high-throughput methods for quantifying amino acids from large maize populations, such as those used for quantitative trait locus mapping, genome-wide association studies, and large mutant populations. Both methods use an ultraperformance liquid chromatography-tandem mass spectrometry instrument to quantify all 20 proteogenic amino acids in a maize tissue in a short run time. A dependable, affordable, and high-throughput method for quantifying amino acids in maize has important implications for assessing kernel quality, yield, and management efficacy, such as fertilizer usage and watering.","PeriodicalId":10496,"journal":{"name":"Cold Spring Harbor protocols","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-12-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142881474","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Examining Cell-Specific Localization in Aedes aegypti Tissues with Fluorescence In Situ Hybridization. 利用荧光原位杂交技术检查埃及伊蚊组织中的特异性细胞定位。

Cold Spring Harbor protocols Pub Date : 2024-12-02 DOI: 10.1101/pdb.prot108280

Farwa Sajadi, Jean-Paul V Paluzzi

引用次数: 0

Filleting and Immunostaining of Larvae to Visualize Drosophila Dendritic Arborization Neuron Dendrite Arbors. 对幼虫进行切片和免疫染色以观察果蝇的树突分化神经元树突节。

Cold Spring Harbor protocols Pub Date : 2024-12-02 DOI: 10.1101/pdb.prot108148

Fangke Xu, Jason Y Tann, Oliver R Wilkes, Adrian W Moore

引用次数: 0

Membrane Protein Fractionation and Analysis through Western Blot in Aedes aegypti Malpighian Tubules. 埃及伊蚊马氏管的膜蛋白分馏和 Western 印迹分析。

Cold Spring Harbor protocols Pub Date : 2024-12-02 DOI: 10.1101/pdb.prot108283

Britney Picinic, Jean-Paul V Paluzzi

引用次数: 0

Mapping Transcript Cell-Specific Localization and Protein Subcellular Localization in the Adult Mosquito Aedes aegypti. 绘制埃及伊蚊成蚊转录本细胞特异性定位和蛋白质亚细胞定位图。

Cold Spring Harbor protocols Pub Date : 2024-12-02 DOI: 10.1101/pdb.top107698

Farwa Sajadi, Salwa Afifi, Britney Picinic, Jean-Paul V Paluzzi

{"title":"Mapping Transcript Cell-Specific Localization and Protein Subcellular Localization in the Adult Mosquito Aedes aegypti.","authors":"Farwa Sajadi, Salwa Afifi, Britney Picinic, Jean-Paul V Paluzzi","doi":"10.1101/pdb.top107698","DOIUrl":"10.1101/pdb.top107698","url":null,"abstract":"This introduction reviews techniques used to examine the distribution and expression of gene transcripts and proteins in a variety of tissues/organs in the medically important global disease vector mosquito, Aedes aegypti Specifically, these methods allow the detection of cell-specific transcript expression by fluorescent in situ hybridization; facilitate immunohistochemical mapping of a protein of interest in whole-mount small tissue/organ samples; examine the subcellular localization of proteins, such as membrane transporters, through sectioning of paraffin-embedded tissue/organ samples; and finally, enable the efficient separation of cytosolic and membrane proteins for western blot analysis without the need for specialized equipment (e.g., ultracentrifuge) in the mosquito Ae. aegypti Such techniques are useful to help answer fundamental questions in mosquito scientific research including (but not limited to) the identification of specific cells in an organ responsible for expressing a receptor of particular interest and necessary for eliciting a response to exogenous signals, including hormones. Moreover, changes in the subcellular localization of specific targets of interest can be assessed both qualitatively and quantitatively, providing insight into transient or long-term physiologically relevant regulation necessary for activity under experimental treatments or varied internal (e.g., development) or external (e.g., environmental stress) factors that might be normally experienced by the organism.","PeriodicalId":10496,"journal":{"name":"Cold Spring Harbor protocols","volume":" ","pages":"pdb.top107698"},"PeriodicalIF":0.0,"publicationDate":"2024-12-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138795556","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Examining Apical or Basolateral Protein Localization in Aedes aegypti Tissues: Cross-Section Immunohistochemistry. 检查埃及伊蚊组织中的顶端或基底侧蛋白定位：横切面免疫组化。

Cold Spring Harbor protocols Pub Date : 2024-12-02 DOI: 10.1101/pdb.prot108282

Farwa Sajadi, Britney Picinic, Salwa Afifi, Jean-Paul V Paluzzi

引用次数: 0

Use of DeTerm for Automated Drosophila Dendrite Arbor Terminal Counts. 使用 DeTerm 自动进行果蝇树突顶端计数。

Cold Spring Harbor protocols Pub Date : 2024-12-02 DOI: 10.1101/pdb.prot108151

Minami Kimura, Jason Y Tann, Oliver R Wilkes, Fangke Xu, Henrik Skibbe, Adrian W Moore

引用次数: 0

Examining Protein Localization in Aedes aegypti Cells, Tissues, and Organs: Whole-Mount Immunohistochemistry. 检查埃及伊蚊细胞、组织和器官中的蛋白质定位：整装免疫组化。

Cold Spring Harbor protocols Pub Date : 2024-12-02 DOI: 10.1101/pdb.prot108281

Salwa Afifi, Farwa Sajadi, Jean-Paul V Paluzzi

{"title":"Examining Protein Localization in Aedes aegypti Cells, Tissues, and Organs: Whole-Mount Immunohistochemistry.","authors":"Salwa Afifi, Farwa Sajadi, Jean-Paul V Paluzzi","doi":"10.1101/pdb.prot108281","DOIUrl":"10.1101/pdb.prot108281","url":null,"abstract":"Immunohistochemistry (IHC) is a powerful technique used for visualizing cellular components and determining the presence and/or location of proteins or other macromolecules in tissue samples. The classical IHC process involves the detection of epitopes using a highly specific primary antibody. This is followed by a secondary antibody that is coupled to a reporter molecule or fluorophore and capable of binding to the primary antibody and allowing for protein immunodetection. Although IHC does not routinely provide quantitative results compared to an enzyme-linked immunoassay or western blotting, it permits the localization of the proteins in intact tissues. This protocol describes an IHC assay for whole-body Aedes aegypti mosquito tissues that is used to detect small proteins, specifically neuropeptide hormones. This method is useful for protein detection in whole-mount preparations; however, cross-section IHC is recommended to determine if a protein is localized in the apical versus basolateral membrane of tissues/organs or to visualize immunological distribution in larger, more complex preparations.","PeriodicalId":10496,"journal":{"name":"Cold Spring Harbor protocols","volume":" ","pages":"pdb.prot108281"},"PeriodicalIF":0.0,"publicationDate":"2024-12-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138795544","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Study of Dendrite Differentiation Using Drosophila Dendritic Arborization Neurons. 利用果蝇树突分化神经元研究树突分化

Cold Spring Harbor protocols Pub Date : 2024-12-02 DOI: 10.1101/pdb.top108146

Jason Y Tann, Fangke Xu, Minami Kimura, Oliver R Wilkes, Li-Foong Yoong, Henrik Skibbe, Adrian W Moore

引用次数: 0

Mounting of Embryos, Larvae, and Pupae for Live Drosophila Dendritic Arborization Neuron Imaging. 为果蝇树突分化神经元活体成像安装胚胎、幼虫和蛹。

Cold Spring Harbor protocols Pub Date : 2024-12-02 DOI: 10.1101/pdb.prot108149

Fangke Xu, Jason Y Tann, Oliver R Wilkes, Minami Kimura, Li-Foong Yoong, Adrian W Moore

引用次数: 0