The Experts below are selected from a list of 128499 Experts worldwide ranked by ideXlab platform
Ming Tsar Chan - One of the best experts on this subject based on the ideXlab platform.
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tape arabidopsis sandwich a simpler arabidopsis protoplast Isolation Method
Plant Methods, 2009Co-Authors: Fu Hui Wu, Shu Chen Shen, Ming Tsar ChanAbstract:Background Protoplasts isolated from leaves are useful materials in plant research. One application, the transient expression of recombinant genes using Arabidopsis mesophyll protoplasts (TEAMP), is currently commonly used for studies of subcellular protein localization, promoter activity, and in vivo protein-protein interactions. This Method requires cutting leaves into very thin slivers to collect mesophyll cell protoplasts, a procedure that often causes cell damage, may yield only a few good protoplasts, and is time consuming. In addition, this protoplast Isolation Method normally requires a large number of leaves derived from plants grown specifically under low-light conditions, which may be a concern when material availability is limited such as with mutant plants, or in large scale experiments.
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tape arabidopsis sandwich a simpler arabidopsis protoplast Isolation Method
Plant Methods, 2009Co-Authors: Shu Chen Shen, Ming Tsar Chan, Lanying Lee, Shu Hong Lee, Chounsea LinAbstract:Protoplasts isolated from leaves are useful materials in plant research. One application, the transient expression of recombinant genes using Arabidopsis mesophyll protoplasts (TEAMP), is currently commonly used for studies of subcellular protein localization, promoter activity, and in vivo protein-protein interactions. This Method requires cutting leaves into very thin slivers to collect mesophyll cell protoplasts, a procedure that often causes cell damage, may yield only a few good protoplasts, and is time consuming. In addition, this protoplast Isolation Method normally requires a large number of leaves derived from plants grown specifically under low-light conditions, which may be a concern when material availability is limited such as with mutant plants, or in large scale experiments. In this report, we present a new procedure that we call the Tape-Arabidopsis Sandwich. This is a simple and fast mesophyll protoplast Isolation Method. Two kinds of tape (Time tape adhered to the upper epidermis and 3 M Magic tape to the lower epidermis) are used to make a "Tape-Arabidopsis Sandwich". The Time tape supports the top side of the leaf during manipulation, while tearing off the 3 M Magic tape allows easy removal of the lower epidermal layer and exposes mesophyll cells to cell wall digesting enzymes when the leaf is later incubated in an enzyme solution. The protoplasts released into solution are collected and washed for further use. For TEAMP, plasmids carrying a gene expression cassette for a fluorescent protein can be successfully delivered into protoplasts isolated from mature leaves grown under optimal conditions. Alternatively, these protoplasts may be used for bimolecular fluorescence complementation (BiFC) to investigate protein-protein interactions in vivo, or for Western blot analysis. A significant advantage of this protocol over the current Method is that it allows the generation of protoplasts in less than 1 hr, and allows TEAMP transfection to be carried out within 2 hr. The protoplasts generated by this new Tape-Arabidopsis Sandwich Method are suitable for the same range of research applications as those that use the current Method, but require less operator skill, equipment and time.
Maire Peters - One of the best experts on this subject based on the ideXlab platform.
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comparison of serum exosome Isolation Methods for microrna profiling
Clinical Biochemistry, 2014Co-Authors: Kadri Rekker, Merli Saare, Anne Mari Roost, Annaliisa Kubo, Natasa Zarovni, Antonio Chiesi, Andres Salumets, Maire PetersAbstract:Abstract Objectives Exosomes are small membrane bound vesicles secreted by most cell types. Exosomes contain various functional proteins, mRNAs and microRNAs (miRNAs) that could be used for diagnostic and therapeutic purposes. Currently, a standard Method for serum exosome Isolation is differential ultracentrifugation, but a search for alternative, less time-consuming and labour extensive exosomal Isolation Method for use in clinical settings is ongoing. The effect of serum exosome Isolation Method on obtained miRNA profile is not yet clear. The aim of this study was to determine to which extent selected exosome Isolation Methods influence the serum exosomal miRNA profile. Design and Methods Exosomes were isolated from blood serum of healthy individuals by ultracentrifugation and ExoQuick Precipitation Methods. The expression profile of 375 miRNAs was determined by real time PCR using Exiqon miRCURY LNA™ microRNA Human panel I assays. Results Although a strong correlation of exosomal miRNA profiles was observed between the two Isolation Methods, distinct clusters of miRNA levels between the used Methods were identified. The detected levels of two miRNAs, miR-92a and miR-486-5p, were significantly influenced by the exosome Isolation Method used. Conclusions Both exosome Isolation Methods are suitable for serum exosomal miRNA profiling. Differences found in miRNA patterns between the two Methods indicate that the observed exosomal miRNA profile is slightly affected by the extracellular vesicle Isolation Method.
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Comparison of serum exosome Isolation Methods for microRNA profiling.
Clinical biochemistry, 2013Co-Authors: Kadri Rekker, Merli Saare, Anne Mari Roost, Annaliisa Kubo, Natasa Zarovni, Antonio Chiesi, Andres Salumets, Maire PetersAbstract:Exosomes are small membrane bound vesicles secreted by most cell types. Exosomes contain various functional proteins, mRNAs and microRNAs (miRNAs) that could be used for diagnostic and therapeutic purposes. Currently, a standard Method for serum exosome Isolation is differential ultracentrifugation, but a search for alternative, less time-consuming and labour extensive exosomal Isolation Method for use in clinical settings is ongoing. The effect of serum exosome Isolation Method on obtained miRNA profile is not yet clear. The aim of this study was to determine to which extent selected exosome Isolation Methods influence the serum exosomal miRNA profile. Exosomes were isolated from blood serum of healthy individuals by ultracentrifugation and ExoQuick Precipitation Methods. The expression profile of 375 miRNAs was determined by real time PCR using Exiqon miRCURY LNA™ microRNA Human panel I assays. Although a strong correlation of exosomal miRNA profiles was observed between the two Isolation Methods, distinct clusters of miRNA levels between the used Methods were identified. The detected levels of two miRNAs, miR-92a and miR-486-5p, were significantly influenced by the exosome Isolation Method used. Both exosome Isolation Methods are suitable for serum exosomal miRNA profiling. Differences found in miRNA patterns between the two Methods indicate that the observed exosomal miRNA profile is slightly affected by the extracellular vesicle Isolation Method. © 2013.
Robert A. Reamer - One of the best experts on this subject based on the ideXlab platform.
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N‐Boc Deprotection and Isolation Method for Water‐Soluble Zwitterionic Compounds.
ChemInform, 2015Co-Authors: Zhijian Liu, Nobuyoshi Yasuda, Michael Simeone, Robert A. ReamerAbstract:An efficient iodotrimethylsilane-mediated deprotection and direct Isolation Method to obtain zwitterionic compounds from the corresponding N-Boc derivatives is developed.
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N-Boc Deprotection and Isolation Method for Water-Soluble Zwitterionic Compounds
The Journal of Organic Chemistry, 2014Co-Authors: Zhijian Liu, Nobuyoshi Yasuda, Michael Simeone, Robert A. ReamerAbstract:A highly efficient TMSI-mediated deprotection and direct Isolation Method to obtain zwitterionic compounds from the corresponding N-Boc derivatives has been developed. This Method has been demonstrated in the final deprotection/Isolation of the β-lactamase inhibitor MK-7655 as a part of its manufacturing process. Further application of this process toward other zwitterionic compounds, such as dipeptides and tripeptides, has been successfully developed. Furthermore, a catalytic version of this transformation has been demonstrated in the presence of BSA or BSTFA.
Shu Chen Shen - One of the best experts on this subject based on the ideXlab platform.
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tape arabidopsis sandwich a simpler arabidopsis protoplast Isolation Method
Plant Methods, 2009Co-Authors: Fu Hui Wu, Shu Chen Shen, Ming Tsar ChanAbstract:Background Protoplasts isolated from leaves are useful materials in plant research. One application, the transient expression of recombinant genes using Arabidopsis mesophyll protoplasts (TEAMP), is currently commonly used for studies of subcellular protein localization, promoter activity, and in vivo protein-protein interactions. This Method requires cutting leaves into very thin slivers to collect mesophyll cell protoplasts, a procedure that often causes cell damage, may yield only a few good protoplasts, and is time consuming. In addition, this protoplast Isolation Method normally requires a large number of leaves derived from plants grown specifically under low-light conditions, which may be a concern when material availability is limited such as with mutant plants, or in large scale experiments.
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tape arabidopsis sandwich a simpler arabidopsis protoplast Isolation Method
Plant Methods, 2009Co-Authors: Shu Chen Shen, Ming Tsar Chan, Lanying Lee, Shu Hong Lee, Chounsea LinAbstract:Protoplasts isolated from leaves are useful materials in plant research. One application, the transient expression of recombinant genes using Arabidopsis mesophyll protoplasts (TEAMP), is currently commonly used for studies of subcellular protein localization, promoter activity, and in vivo protein-protein interactions. This Method requires cutting leaves into very thin slivers to collect mesophyll cell protoplasts, a procedure that often causes cell damage, may yield only a few good protoplasts, and is time consuming. In addition, this protoplast Isolation Method normally requires a large number of leaves derived from plants grown specifically under low-light conditions, which may be a concern when material availability is limited such as with mutant plants, or in large scale experiments. In this report, we present a new procedure that we call the Tape-Arabidopsis Sandwich. This is a simple and fast mesophyll protoplast Isolation Method. Two kinds of tape (Time tape adhered to the upper epidermis and 3 M Magic tape to the lower epidermis) are used to make a "Tape-Arabidopsis Sandwich". The Time tape supports the top side of the leaf during manipulation, while tearing off the 3 M Magic tape allows easy removal of the lower epidermal layer and exposes mesophyll cells to cell wall digesting enzymes when the leaf is later incubated in an enzyme solution. The protoplasts released into solution are collected and washed for further use. For TEAMP, plasmids carrying a gene expression cassette for a fluorescent protein can be successfully delivered into protoplasts isolated from mature leaves grown under optimal conditions. Alternatively, these protoplasts may be used for bimolecular fluorescence complementation (BiFC) to investigate protein-protein interactions in vivo, or for Western blot analysis. A significant advantage of this protocol over the current Method is that it allows the generation of protoplasts in less than 1 hr, and allows TEAMP transfection to be carried out within 2 hr. The protoplasts generated by this new Tape-Arabidopsis Sandwich Method are suitable for the same range of research applications as those that use the current Method, but require less operator skill, equipment and time.
Chounsea Lin - One of the best experts on this subject based on the ideXlab platform.
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tape arabidopsis sandwich a simpler arabidopsis protoplast Isolation Method
Plant Methods, 2009Co-Authors: Shu Chen Shen, Ming Tsar Chan, Lanying Lee, Shu Hong Lee, Chounsea LinAbstract:Protoplasts isolated from leaves are useful materials in plant research. One application, the transient expression of recombinant genes using Arabidopsis mesophyll protoplasts (TEAMP), is currently commonly used for studies of subcellular protein localization, promoter activity, and in vivo protein-protein interactions. This Method requires cutting leaves into very thin slivers to collect mesophyll cell protoplasts, a procedure that often causes cell damage, may yield only a few good protoplasts, and is time consuming. In addition, this protoplast Isolation Method normally requires a large number of leaves derived from plants grown specifically under low-light conditions, which may be a concern when material availability is limited such as with mutant plants, or in large scale experiments. In this report, we present a new procedure that we call the Tape-Arabidopsis Sandwich. This is a simple and fast mesophyll protoplast Isolation Method. Two kinds of tape (Time tape adhered to the upper epidermis and 3 M Magic tape to the lower epidermis) are used to make a "Tape-Arabidopsis Sandwich". The Time tape supports the top side of the leaf during manipulation, while tearing off the 3 M Magic tape allows easy removal of the lower epidermal layer and exposes mesophyll cells to cell wall digesting enzymes when the leaf is later incubated in an enzyme solution. The protoplasts released into solution are collected and washed for further use. For TEAMP, plasmids carrying a gene expression cassette for a fluorescent protein can be successfully delivered into protoplasts isolated from mature leaves grown under optimal conditions. Alternatively, these protoplasts may be used for bimolecular fluorescence complementation (BiFC) to investigate protein-protein interactions in vivo, or for Western blot analysis. A significant advantage of this protocol over the current Method is that it allows the generation of protoplasts in less than 1 hr, and allows TEAMP transfection to be carried out within 2 hr. The protoplasts generated by this new Tape-Arabidopsis Sandwich Method are suitable for the same range of research applications as those that use the current Method, but require less operator skill, equipment and time.
