The Experts below are selected from a list of 156 Experts worldwide ranked by ideXlab platform
Jeremy Nathans - One of the best experts on this subject based on the ideXlab platform.
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Identification and characterization of all-trans-retinol dehydrogenase from photoreceptor outer segments, the visual cycle enzyme that reduces all-trans-retinal to all-trans-retinol.
The Journal of biological chemistry, 2000Co-Authors: Amir Rattner, Philip M. Smallwood, Jeremy NathansAbstract:Retinol dehydrogenase (RDH), the enzyme that catalyzes the reduction of all-trans-retinal to all-trans-retinol within the photoreceptor outer segment, was the first visual cycle enzymatic activity to be identified. Previous work has shown that this enzyme utilizes NADPH, shows a marked preference for all-trans-retinal over 11-cis-retinal, and is tightly associated with the outer segment membrane. This paper reports the identification of a novel member of the short chain dehydrogenase/reductase family, photoreceptor RDH (prRDH), using subtraction and normalization of retina cDNA, high throughput sequencing, and data base homology searches to detect retina-specific genes. Bovine and human prRDH are highly homologous and are most closely related to 17-beta-hydroxysteroid dehydrogenase 1. The enzymatic properties of recombinant bovine prRDH closely match those previously reported for RDH activity in crude bovine rod outer segment preparations. In situ hybridization and RNA Blotting show that the PRRDH gene is expressed specifically in photoreceptor cells, and Protein Blotting and immunocytochemistry show that prRDH localizes exclusively to both rod and cone outer segments and that prRDH is tightly associated with outer segment membranes. Taken together, these data indicate that prRDH is the enzyme responsible for the reduction of all-trans-retinal to all-trans-retinol within the photoreceptor outer segment.
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Identification and characterization of all-trans-retinol dehydrogenase from photoreceptor outer segments, the visual cycle enzyme that reduces all-trans-retinal to all-trans-retinol.
Journal of Biological Chemistry, 2000Co-Authors: Amir Rattner, Philip M. Smallwood, Jeremy NathansAbstract:Abstract Retinol dehydrogenase (RDH), the enzyme that catalyzes the reduction of all-trans-retinal to all-trans-retinol within the photoreceptor outer segment, was the first visual cycle enzymatic activity to be identified. Previous work has shown that this enzyme utilizes NADPH, shows a marked preference for all-trans-retinal over 11-cis-retinal, and is tightly associated with the outer segment membrane. This paper reports the identification of a novel member of the short chain dehydrogenase/reductase family, photoreceptor RDH (prRDH), using subtraction and normalization of retina cDNA, high throughput sequencing, and data base homology searches to detect retina-specific genes. Bovine and human prRDH are highly homologous and are most closely related to 17-β-hydroxysteroid dehydrogenase 1. The enzymatic properties of recombinant bovine prRDH closely match those previously reported for RDH activity in crude bovine rod outer segment preparations. In situ hybridization and RNA Blotting show that the PRRDH gene is expressed specifically in photoreceptor cells, and Protein Blotting and immunocytochemistry show that prRDH localizes exclusively to both rod and cone outer segments and that prRDH is tightly associated with outer segment membranes. Taken together, these data indicate that prRDH is the enzyme responsible for the reduction of all-trans-retinal to all-trans-retinol within the photoreceptor outer segment.
Amir Rattner - One of the best experts on this subject based on the ideXlab platform.
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Identification and characterization of all-trans-retinol dehydrogenase from photoreceptor outer segments, the visual cycle enzyme that reduces all-trans-retinal to all-trans-retinol.
The Journal of biological chemistry, 2000Co-Authors: Amir Rattner, Philip M. Smallwood, Jeremy NathansAbstract:Retinol dehydrogenase (RDH), the enzyme that catalyzes the reduction of all-trans-retinal to all-trans-retinol within the photoreceptor outer segment, was the first visual cycle enzymatic activity to be identified. Previous work has shown that this enzyme utilizes NADPH, shows a marked preference for all-trans-retinal over 11-cis-retinal, and is tightly associated with the outer segment membrane. This paper reports the identification of a novel member of the short chain dehydrogenase/reductase family, photoreceptor RDH (prRDH), using subtraction and normalization of retina cDNA, high throughput sequencing, and data base homology searches to detect retina-specific genes. Bovine and human prRDH are highly homologous and are most closely related to 17-beta-hydroxysteroid dehydrogenase 1. The enzymatic properties of recombinant bovine prRDH closely match those previously reported for RDH activity in crude bovine rod outer segment preparations. In situ hybridization and RNA Blotting show that the PRRDH gene is expressed specifically in photoreceptor cells, and Protein Blotting and immunocytochemistry show that prRDH localizes exclusively to both rod and cone outer segments and that prRDH is tightly associated with outer segment membranes. Taken together, these data indicate that prRDH is the enzyme responsible for the reduction of all-trans-retinal to all-trans-retinol within the photoreceptor outer segment.
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Identification and characterization of all-trans-retinol dehydrogenase from photoreceptor outer segments, the visual cycle enzyme that reduces all-trans-retinal to all-trans-retinol.
Journal of Biological Chemistry, 2000Co-Authors: Amir Rattner, Philip M. Smallwood, Jeremy NathansAbstract:Abstract Retinol dehydrogenase (RDH), the enzyme that catalyzes the reduction of all-trans-retinal to all-trans-retinol within the photoreceptor outer segment, was the first visual cycle enzymatic activity to be identified. Previous work has shown that this enzyme utilizes NADPH, shows a marked preference for all-trans-retinal over 11-cis-retinal, and is tightly associated with the outer segment membrane. This paper reports the identification of a novel member of the short chain dehydrogenase/reductase family, photoreceptor RDH (prRDH), using subtraction and normalization of retina cDNA, high throughput sequencing, and data base homology searches to detect retina-specific genes. Bovine and human prRDH are highly homologous and are most closely related to 17-β-hydroxysteroid dehydrogenase 1. The enzymatic properties of recombinant bovine prRDH closely match those previously reported for RDH activity in crude bovine rod outer segment preparations. In situ hybridization and RNA Blotting show that the PRRDH gene is expressed specifically in photoreceptor cells, and Protein Blotting and immunocytochemistry show that prRDH localizes exclusively to both rod and cone outer segments and that prRDH is tightly associated with outer segment membranes. Taken together, these data indicate that prRDH is the enzyme responsible for the reduction of all-trans-retinal to all-trans-retinol within the photoreceptor outer segment.
Philip M. Smallwood - One of the best experts on this subject based on the ideXlab platform.
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Identification and characterization of all-trans-retinol dehydrogenase from photoreceptor outer segments, the visual cycle enzyme that reduces all-trans-retinal to all-trans-retinol.
The Journal of biological chemistry, 2000Co-Authors: Amir Rattner, Philip M. Smallwood, Jeremy NathansAbstract:Retinol dehydrogenase (RDH), the enzyme that catalyzes the reduction of all-trans-retinal to all-trans-retinol within the photoreceptor outer segment, was the first visual cycle enzymatic activity to be identified. Previous work has shown that this enzyme utilizes NADPH, shows a marked preference for all-trans-retinal over 11-cis-retinal, and is tightly associated with the outer segment membrane. This paper reports the identification of a novel member of the short chain dehydrogenase/reductase family, photoreceptor RDH (prRDH), using subtraction and normalization of retina cDNA, high throughput sequencing, and data base homology searches to detect retina-specific genes. Bovine and human prRDH are highly homologous and are most closely related to 17-beta-hydroxysteroid dehydrogenase 1. The enzymatic properties of recombinant bovine prRDH closely match those previously reported for RDH activity in crude bovine rod outer segment preparations. In situ hybridization and RNA Blotting show that the PRRDH gene is expressed specifically in photoreceptor cells, and Protein Blotting and immunocytochemistry show that prRDH localizes exclusively to both rod and cone outer segments and that prRDH is tightly associated with outer segment membranes. Taken together, these data indicate that prRDH is the enzyme responsible for the reduction of all-trans-retinal to all-trans-retinol within the photoreceptor outer segment.
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Identification and characterization of all-trans-retinol dehydrogenase from photoreceptor outer segments, the visual cycle enzyme that reduces all-trans-retinal to all-trans-retinol.
Journal of Biological Chemistry, 2000Co-Authors: Amir Rattner, Philip M. Smallwood, Jeremy NathansAbstract:Abstract Retinol dehydrogenase (RDH), the enzyme that catalyzes the reduction of all-trans-retinal to all-trans-retinol within the photoreceptor outer segment, was the first visual cycle enzymatic activity to be identified. Previous work has shown that this enzyme utilizes NADPH, shows a marked preference for all-trans-retinal over 11-cis-retinal, and is tightly associated with the outer segment membrane. This paper reports the identification of a novel member of the short chain dehydrogenase/reductase family, photoreceptor RDH (prRDH), using subtraction and normalization of retina cDNA, high throughput sequencing, and data base homology searches to detect retina-specific genes. Bovine and human prRDH are highly homologous and are most closely related to 17-β-hydroxysteroid dehydrogenase 1. The enzymatic properties of recombinant bovine prRDH closely match those previously reported for RDH activity in crude bovine rod outer segment preparations. In situ hybridization and RNA Blotting show that the PRRDH gene is expressed specifically in photoreceptor cells, and Protein Blotting and immunocytochemistry show that prRDH localizes exclusively to both rod and cone outer segments and that prRDH is tightly associated with outer segment membranes. Taken together, these data indicate that prRDH is the enzyme responsible for the reduction of all-trans-retinal to all-trans-retinol within the photoreceptor outer segment.
Jiri Moos - One of the best experts on this subject based on the ideXlab platform.
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DETECTION OF GELATINOLYTIC ENZYME ACTIVITIES AFTER SODIUM DODECYL SULFATE-ELECTROPHORESIS AND Protein Blotting
Electrophoresis, 1991Co-Authors: Jiri MoosAbstract:Visualization of proteases with gelatinolytic activity in sodium dodecyl sulfate gels is described. After conventional sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Proteins are transferred onto nitrocellulose membranes preincubated with 0.3% gelatine. During the Protein electrotransfer, the proteases are renaturated and their enzymatic activity is restored. After nonspecific Protein staining, bands with proteolytic activity appear as white areas on a dark background.
R. Hal Scofield - One of the best experts on this subject based on the ideXlab platform.
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Validating Antibody Specificities for Immunohistochemistry by Protein Blotting Methods.
Methods in molecular biology (Clifton N.J.), 2017Co-Authors: Biji T. Kurien, R. Hal ScofieldAbstract:ImmunoBlotting has been used in conjunction with other important antibody based detection methods like enzyme linked immunosorbent assay and immunohistochemistry to provide confirmation of results both in research and diagnostic testing. Specificity of antibodies employed for immunohistochemical studies is of critical importance and therefore the use of western Blotting is imperative to address specificity of antibodies. In spite of its overall simplicity, western Blotting or Protein Blotting is a powerful procedure for immunodetection of Proteins, especially those that are of low abundance, following electrophoretic separation. The usefulness of this procedure stems from its ability to provide simultaneous resolution of multiple immunogenic antigens within a sample for detection by specific antibodies. Protein Blotting has evolved greatly over the last few decades and researchers have a variety of ways and means to carry out this procedure to validate antibodies for immunohistochemistry.
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Other notable Protein Blotting methods: a brief review.
Methods in molecular biology (Clifton N.J.), 2015Co-Authors: Biji T. Kurien, R. Hal ScofieldAbstract:Proteins have been transferred from the gel to the membrane by a variety of methods. These include vacuum Blotting, centrifuge Blotting, electroBlotting of Proteins to Teflon tape and membranes for N- and C-terminal sequence analysis, multiple tissue Blotting, a two-step transfer of low- and high-molecular-weight Proteins, acid electroBlotting onto activated glass, membrane-array method for the detection of human intestinal bacteria in fecal samples, Protein microarray using a new black cellulose nitrate support, electrotransfer using square wave alternating voltage for enhanced Protein recovery, polyethylene glycol-mediated significant enhancement of the immunoBlotting transfer, parallel Protein chemical processing before and during western blot and the molecular scanner concept, electronic western blot of matrix-assisted laser desorption/ionization mass spectrometric-identified polypeptides from parallel processed gel-separated Proteins, semidry electroBlotting of peptides and Proteins from acid-urea polyacrylamide gels, transfer of silver-stained Proteins from polyacrylamide gels to polyvinylidene difluoride (PVDF) membranes, and the display of K(+) channel Proteins on a solid nitrocellulose support for assaying toxin binding. The quantification of Proteins bound to PVDF membranes by elution of CBB, clarification of immunoblots on PVDF for transmission densitometry, gold coating of nonconductive membranes before matrix-assisted laser desorption/ionization tandem mass spectrometric analysis to prevent charging effect for analysis of peptides from PVDF membranes, and a simple method for coating native polysaccharides onto nitrocellulose are some of the methods involving either the manipulation of membranes with transferred Proteins or just a passive transfer of antigens to membranes. All these methods are briefly reviewed in this chapter.
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Introduction to Protein Blotting.
Methods in molecular biology (Clifton N.J.), 2009Co-Authors: Biji T. Kurien, R. Hal ScofieldAbstract:Protein Blotting is a powerful and important procedure for the immunodetection of Proteins following electrophoresis, particularly Proteins that are of low abundance. Since the inception of the protocol for Protein transfer from an electrophoresed gel to a membrane in 1979, Protein Blotting has evolved greatly. The scientific community is now confronted with a variety of ways and means to carry out this transfer.
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A brief review of other notable Protein Blotting methods.
Methods in molecular biology (Clifton N.J.), 2009Co-Authors: Biji T. Kurien, R. Hal ScofieldAbstract:A plethora of methods have been used for transferring Proteins from the gel to the membrane. These include centrifuge Blotting, electroBlotting of Proteins to Teflon tape and membranes for N- and C-terminal sequence analysis, multiple tissue Blotting, a two-step transfer of low and high molecular weight Proteins, Blotting of Coomassie Brilliant Blue (CBB)-stained Proteins from polyacrylamide gels to transparencies, acid electroBlotting onto activated glass, membrane-array method for the detection of human intestinal bacteria in fecal samples, Protein microarray using a new black cellulose nitrate support, electrotransfer using square wave alternating voltage for enhanced Protein recovery, polyethylene glycol-mediated significant enhancement of the immunoBlotting transfer, parallel Protein chemical processing before and during western blot and the molecular scanner concept, electronic western blot of matrix-assisted laser desorption/ionization (MALDI) mass spectrometry-identified polypeptides from parallel processed gel-separated Proteins, semidry electroBlotting of peptides and Proteins from acid-urea polyacrylamide gels, transfer of silver-stained Proteins from polyacrylamide gels to polyvinylidene difluoride (PVDF) membranes, and the display of K(+) channel Proteins on a solid nitrocellulose support for assaying toxin binding. The quantification of Proteins bound to PVDF membranes by elution of CBB, clarification of immunoblots on PVDF for transmission densitometry, gold coating of nonconductive membranes before MALDI tandem mass spectrometric analysis to prevent charging effect for analysis of peptides from PVDF membranes, and a simple method for coating native polysaccharides onto nitrocellulose are some of the methods involving either the manipulation of membranes with transferred Proteins or just a passive transfer of antigens to membranes. All these methods are briefly reviewed in this chapter.
