The Experts below are selected from a list of 204 Experts worldwide ranked by ideXlab platform

Thierry Rabilloud - One of the best experts on this subject based on the ideXlab platform.

  • Silver Staining of 2D Electrophoresis Gels.
    Methods in Molecular Biology, 2012
    Co-Authors: Thierry Rabilloud
    Abstract:

    Silver Staining is used to detect proteins after electrophoretic separation on polyacrylamide gels. It -combines excellent sensitivity (in the low nanogram range) with the use of very simple and cheap equipment and chemicals. For its use in proteomics, two important additional features must be considered, compatibility with mass spectrometry and quantitative response. Both features are discussed in this chapter, and optimized Silver Staining protocols are proposed.

  • Silver Staining of Proteins in 2DE Gels.
    Methods in Molecular Biology, 2009
    Co-Authors: Cécile Lelong, Sylvie Luche, Mireille Chevallet, Thierry Rabilloud
    Abstract:

    Silver Staining detects proteins after electrophoretic separation on polyacrylamide gels. Its main positive features are its excellent sensitivity (in the low nanogram range) and the use of very simple and cheap equipment and chemicals. The sequential phases of Silver Staining are protein fixation, then sensitization, then Silver impregnation, and finally image development. Several variants of Silver Staining are described here, which can be completed in a time range from 2 h to 1 day after the end of the electrophoretic separation. Once completed, the stain is stable for several weeks.

  • Sweet Silver: A formaldehyde-free Silver Staining using aldoses as developing agents, with enhanced compatibility with mass spectrometry.
    Proteomics, 2008
    Co-Authors: Mireille Chevallet, Sylvie Luche, Alain Van Dorsselaer, Hélène Diemer, Jean-marc Strub, Thierry Rabilloud
    Abstract:

    Protein detection methods after electrophoresis have to be sensitive, homogeneous, and not to impair downstream analysis of proteins by MS. Speed, low cost, and user friendliness are also favored features. Silver Staining combines many of these features, but its compatibility with MS is limited. We describe here, a new variant of Silver Staining that is completely formaldehyde-free. Reducing sugars in alkaline borate buffer are used as developers. While keeping the benefits of Silver Staining, this method is shown to afford a much better performance in terms of compatibility with MS, both in PMF by MALDI and in LC/ESI/MS/MS.

  • Silver Staining of proteins in polyacrylamide gels
    Nature Protocols, 2006
    Co-Authors: Mireille Chevallet, Sylvie Luche, Thierry Rabilloud
    Abstract:

    Silver Staining is used to detect proteins after electrophoretic separation on polyacrylamide gels. It combines excellent sensitivity (in the low nanogram range) with the use of very simple and cheap equipment and chemicals. It is compatible with downstream processing, such as mass spectrometry analysis after protein digestion. The sequential phases of Silver Staining are protein fixation, then sensitization, then Silver impregnation and finally image development. Several variants of Silver Staining are described here, which can be completed in a time range from 2 h to 1 d after the end of the electrophoretic separation. Once completed, the stain is stable for several weeks.

  • About the mechanism of interference of Silver Staining with peptide mass spectrometry.
    Proteomics, 2004
    Co-Authors: Sophie Richert, Sylvie Luche, Mireille Chevallet, Alain Van Dorsselaer, Emmanuelle Leize-wagner, Thierry Rabilloud
    Abstract:

    The mechanism by which Silver Staining of proteins in polyacrylamide gels interferes with mass spectrometry of peptides produced by proteolysis has been investigated. It was demonstrated that this interference increases with time between Silver Staining and gel processing, although the Silver image is constant. This suggested an important role of the formaldehyde used in Silver Staining development in this interference process. Consequently, a formaldehyde-free Staining protocol has been devised, using carbohydrazide as the developing agent. This protocol showed much increased peptide coverage and retained the sensitivity of Silver Staining. These results were however obtained at the expense of an increased background in the stained gels and of a reduced Staining homogeneity.

Michael Bachmann - One of the best experts on this subject based on the ideXlab platform.

  • Silver Staining Techniques of Polyacrylamide Gels
    Methods in molecular biology (Clifton N.J.), 2018
    Co-Authors: Nicole Berndt, Claudia Arndt, Stefanie Koristka, Ralf Bergmann, Michael Bachmann
    Abstract:

    After SDS-polyacrylamide gel electrophoresis the separated proteins have to be visualized by Staining the gel. The same is true after transfer of separated proteins to a blotting membrane in order to verify an efficient transfer and to visualize the amount of protein(s) remaining in the gel. Several different Staining techniques exist for Staining of proteins in SDS-polyacrylamide gels. The sensitivity of these Staining procedures are different, also the expenditure of time and other aspects. Still, Silver Staining is among the most sensitive and reliable Staining technique. Because this technique was developed in the 1970s, a huge number of variations exist. Here, we will provide three variations, which are robust and easy to perform.

  • Silver Staining techniques of polyacrylamide gels.
    Methods in molecular biology (Clifton N.J.), 2012
    Co-Authors: Holger Bartsch, Claudia Arndt, Stefanie Koristka, Marc Cartellieri, Michael Bachmann
    Abstract:

    Although the main application for polyacrylamide gels is the separation and subsequent blotting of proteins for immunodetection, there are tasks that need Staining of proteins in the polyacrylamide gel. Several different Staining techniques exist for protein Staining in SDS gels that differ in their sensitivity, their expenditure of time, and other aspects. Still, Silver Staining is the most sensitive and reliable Staining technique. Because this technique was developed in the 1970s, a huge number of variations exist. Therefore, we will provide herein three methods, which are robust and easy to perform.

Mireille Chevallet - One of the best experts on this subject based on the ideXlab platform.

  • Silver Staining of Proteins in 2DE Gels.
    Methods in Molecular Biology, 2009
    Co-Authors: Cécile Lelong, Sylvie Luche, Mireille Chevallet, Thierry Rabilloud
    Abstract:

    Silver Staining detects proteins after electrophoretic separation on polyacrylamide gels. Its main positive features are its excellent sensitivity (in the low nanogram range) and the use of very simple and cheap equipment and chemicals. The sequential phases of Silver Staining are protein fixation, then sensitization, then Silver impregnation, and finally image development. Several variants of Silver Staining are described here, which can be completed in a time range from 2 h to 1 day after the end of the electrophoretic separation. Once completed, the stain is stable for several weeks.

  • Sweet Silver: A formaldehyde-free Silver Staining using aldoses as developing agents, with enhanced compatibility with mass spectrometry.
    Proteomics, 2008
    Co-Authors: Mireille Chevallet, Sylvie Luche, Alain Van Dorsselaer, Hélène Diemer, Jean-marc Strub, Thierry Rabilloud
    Abstract:

    Protein detection methods after electrophoresis have to be sensitive, homogeneous, and not to impair downstream analysis of proteins by MS. Speed, low cost, and user friendliness are also favored features. Silver Staining combines many of these features, but its compatibility with MS is limited. We describe here, a new variant of Silver Staining that is completely formaldehyde-free. Reducing sugars in alkaline borate buffer are used as developers. While keeping the benefits of Silver Staining, this method is shown to afford a much better performance in terms of compatibility with MS, both in PMF by MALDI and in LC/ESI/MS/MS.

  • Silver Staining of proteins in polyacrylamide gels
    Nature Protocols, 2006
    Co-Authors: Mireille Chevallet, Sylvie Luche, Thierry Rabilloud
    Abstract:

    Silver Staining is used to detect proteins after electrophoretic separation on polyacrylamide gels. It combines excellent sensitivity (in the low nanogram range) with the use of very simple and cheap equipment and chemicals. It is compatible with downstream processing, such as mass spectrometry analysis after protein digestion. The sequential phases of Silver Staining are protein fixation, then sensitization, then Silver impregnation and finally image development. Several variants of Silver Staining are described here, which can be completed in a time range from 2 h to 1 d after the end of the electrophoretic separation. Once completed, the stain is stable for several weeks.

  • About the mechanism of interference of Silver Staining with peptide mass spectrometry.
    Proteomics, 2004
    Co-Authors: Sophie Richert, Sylvie Luche, Mireille Chevallet, Alain Van Dorsselaer, Emmanuelle Leize-wagner, Thierry Rabilloud
    Abstract:

    The mechanism by which Silver Staining of proteins in polyacrylamide gels interferes with mass spectrometry of peptides produced by proteolysis has been investigated. It was demonstrated that this interference increases with time between Silver Staining and gel processing, although the Silver image is constant. This suggested an important role of the formaldehyde used in Silver Staining development in this interference process. Consequently, a formaldehyde-free Staining protocol has been devised, using carbohydrazide as the developing agent. This protocol showed much increased peptide coverage and retained the sensitivity of Silver Staining. These results were however obtained at the expense of an increased background in the stained gels and of a reduced Staining homogeneity.

Peter M Gresshoff - One of the best experts on this subject based on the ideXlab platform.

  • Silver Staining DNA in polyacrylamide gels
    Nature Protocols, 2007
    Co-Authors: Brant J. Bassam, Peter M Gresshoff
    Abstract:

    This protocol describes a simple Silver Staining method used to visualize DNA fragments and other organic molecules with unsurpassed detail following traditional polyacrylamide gel electrophoresis (PAGE). Sensitivity rivals radioisotopic methods and DNA in the picogram range can be reliably detected. The described protocol is fast (∼1 h) and is implemented using readily available chemicals and materials. To achieve the sensitivity and visual clarity expected, quality reagents and clean handling are important. The updated protocol described here is based on the widely used method of Bassam et al . (1991), but provides improved image contrast and less risk of Staining artefacts.

  • Silver Staining dna in polyacrylamide gels
    Nature Protocols, 2007
    Co-Authors: Brant J. Bassam, Peter M Gresshoff
    Abstract:

    This protocol describes a simple Silver Staining method used to visualize DNA fragments and other organic molecules with unsurpassed detail following traditional polyacrylamide gel electrophoresis (PAGE). Sensitivity rivals radioisotopic methods and DNA in the picogram range can be reliably detected. The described protocol is fast (approximately 1 h) and is implemented using readily available chemicals and materials. To achieve the sensitivity and visual clarity expected, quality reagents and clean handling are important. The updated protocol described here is based on the widely used method of Bassam et al. (1991), but provides improved image contrast and less risk of Staining artefacts.

Sylvie Luche - One of the best experts on this subject based on the ideXlab platform.

  • Silver Staining of Proteins in 2DE Gels.
    Methods in Molecular Biology, 2009
    Co-Authors: Cécile Lelong, Sylvie Luche, Mireille Chevallet, Thierry Rabilloud
    Abstract:

    Silver Staining detects proteins after electrophoretic separation on polyacrylamide gels. Its main positive features are its excellent sensitivity (in the low nanogram range) and the use of very simple and cheap equipment and chemicals. The sequential phases of Silver Staining are protein fixation, then sensitization, then Silver impregnation, and finally image development. Several variants of Silver Staining are described here, which can be completed in a time range from 2 h to 1 day after the end of the electrophoretic separation. Once completed, the stain is stable for several weeks.

  • Sweet Silver: A formaldehyde-free Silver Staining using aldoses as developing agents, with enhanced compatibility with mass spectrometry.
    Proteomics, 2008
    Co-Authors: Mireille Chevallet, Sylvie Luche, Alain Van Dorsselaer, Hélène Diemer, Jean-marc Strub, Thierry Rabilloud
    Abstract:

    Protein detection methods after electrophoresis have to be sensitive, homogeneous, and not to impair downstream analysis of proteins by MS. Speed, low cost, and user friendliness are also favored features. Silver Staining combines many of these features, but its compatibility with MS is limited. We describe here, a new variant of Silver Staining that is completely formaldehyde-free. Reducing sugars in alkaline borate buffer are used as developers. While keeping the benefits of Silver Staining, this method is shown to afford a much better performance in terms of compatibility with MS, both in PMF by MALDI and in LC/ESI/MS/MS.

  • Silver Staining of proteins in polyacrylamide gels
    Nature Protocols, 2006
    Co-Authors: Mireille Chevallet, Sylvie Luche, Thierry Rabilloud
    Abstract:

    Silver Staining is used to detect proteins after electrophoretic separation on polyacrylamide gels. It combines excellent sensitivity (in the low nanogram range) with the use of very simple and cheap equipment and chemicals. It is compatible with downstream processing, such as mass spectrometry analysis after protein digestion. The sequential phases of Silver Staining are protein fixation, then sensitization, then Silver impregnation and finally image development. Several variants of Silver Staining are described here, which can be completed in a time range from 2 h to 1 d after the end of the electrophoretic separation. Once completed, the stain is stable for several weeks.

  • About the mechanism of interference of Silver Staining with peptide mass spectrometry.
    Proteomics, 2004
    Co-Authors: Sophie Richert, Sylvie Luche, Mireille Chevallet, Alain Van Dorsselaer, Emmanuelle Leize-wagner, Thierry Rabilloud
    Abstract:

    The mechanism by which Silver Staining of proteins in polyacrylamide gels interferes with mass spectrometry of peptides produced by proteolysis has been investigated. It was demonstrated that this interference increases with time between Silver Staining and gel processing, although the Silver image is constant. This suggested an important role of the formaldehyde used in Silver Staining development in this interference process. Consequently, a formaldehyde-free Staining protocol has been devised, using carbohydrazide as the developing agent. This protocol showed much increased peptide coverage and retained the sensitivity of Silver Staining. These results were however obtained at the expense of an increased background in the stained gels and of a reduced Staining homogeneity.