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  • 1
    Call number: QH506:60/536
    Keywords: Blotting, Western / methods ; Proteins / analysis
    Pages: xiii, 588 p. : ill.
    ISBN: 9781934115732
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  • 2
    Keywords: Life sciences ; Proteins ; Life sciences ; Protein Science ; Springer eBooks
    Description / Table of Contents: Protein Stains and Applications -- The Roles of Acetic Acid and Methanol during Fixing and Staining Proteins in a SDS Polyacrylamide Electrophoresis Gel -- Multicolored Prestained Standard Protein Marker Generation using a Variety of Remazol Dyes for Easy Visualization of Protein Bands during SDS-PAGE -- Coomassie-Brilliant Blue Staining of Polyacrylamide Gels -- A Simple, Time-Saving Dye Staining of Proteins in Sodium Dodecyl Sulfate Polyacrylamide Gel using Coomassie Blue -- Application of Heat to Quickly Stain and Destain Proteins Stained with Coomassie Blue -- Silver Staining Techniques of Polyacrylamide Gels -- Counterion Dye Staining of Proteins in One- and Two-Dimensional Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis and Tryptic Gel Digestion of Stained Protein for Mass Spectrometry -- Detection of Phosphoproteins in Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis using 8-Quinolinol Stain -- Microwave Assisted Protein Staining, De-Staining and In-Gel/In-Solution Digestion of Proteins -- Fluorescent Staining of Gels -- A Single-Step Simultaneous Protein Staining Procedure for Polyacrylamide Gels and Nitrocellulose Membranes by Alta during Western Blot Analysis -- TEMED Enhanced Photoluminescent Imaging of Human Serum Proteins by Quantum Dots after PAGE -- Detection of Glycoproteins in Polyacrylamide Gels using Pro-Q Emerald 300 Dye, A Fluorescent Periodate Schiff-Base Stain -- Curcumin/Turmeric as an Environment-Friendly Protein Gel Stain -- Detection of Multiple Enzymes in Fermentation Broth using Single PAGE Analysis -- Revisit of Imidazole-Zinc Reverse Stain for Protein Polyacrylamide Gel Electrophoresis -- A One-Step Staining Protocol for In-Gel Fluorescent Visualization of Proteins -- Tem Minute stain to Detect Proteins in Polyacrylamide Electrophoresis Gels with Direct red 81 and Amido Black -- In-Gel Protein Phosphatase Assay using Fluorogenic Substrates -- Detection of Proteins in Polyacrylamide Gels via Pre-Labeling by Isatoic Anhydride -- Fluorescent Protein Visualization Immediately after Gel Electrophoresis using an In-Gel Trichloroethanol Photoreaction with Tryptophan -- Direct Immunodetection of Antigens within the Precast Polyacrylamide Gel -- Zymographic Determination of Intrinsic Specific Activity of Oxidases in Presence of Interfering Proteins -- A Simple Method for Detecting Phosphorylation of Proteins by using ZN2+ -Phos-tag SDS-PAGE at Neutral pH -- Principle and Method of Silver Staining of Proteins Separated by Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis -- Heat/Pressure Treatment with Detergents Significantly Increases Curcumin Solubility and Stability: Its Use as an Environment-Friendly Protein Gel Stain -- Fungal Laccase Efficiently Destains Coomassie Brilliant Blue-R-250 Stained Polyacrylamide Gels -- Destaining Coomassie Brilliant Blue-Stained Sodium Dodecyl Sulfate Polyacrylamide Protein Gels using a Household Detergent -- Paper Adsorbents Remove Coomassie Blue from Gel Destain and used Gel Stain in an Environment-Friendly Manner -- Gel Drying Methods -- Stained Gels can be Stored for Several Months in Non-Sealed Polyethylene Bags -- Radiolabeling and Analysis of Labeled Proteins
    Abstract: This volume expands upon the collection of techniques published in Protein Electrophoresis: Methods and Protocols (2012) with more practical and reproducible methods to study protein gel detection and imaging. The chapters in this book cover topics such as coomassie-brilliant blue staining of polyacrylamide gels; silver staining techniques; microwave assisted protein staining, de-staining, and in-solution digestion of proteins; curumin and turmeric as an environment-friendly protein gel stain; in-gel protein phosphotase assay using fluorogenic substrates; destaining with fungal laccase; and radiolabeling and analysis of labeled proteins. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and practical, Protein Gel Detection and Imaging: Methods and Protocols is a valuable resource for expert and novice scientists and researchers who are interested in learning and experimenting with this field
    Pages: XIII, 289 p. 39 illus., 35 illus. in color. : online resource.
    ISBN: 9781493987450
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  • 3
    Keywords: Biochemistry ; Protein Science ; Springer eBooks
    Description / Table of Contents: How It All Began: A Personal History of Gel-Electrophoresis -- Introduction to Protein Electrophoresis -- Concentrating Proteins by Salt, Polyethylene Glycol, Solvent, SDS Precipitation, Three Phase Partitioning, Dialysis, Centrifugation, Ultrafiltration, Lyophilisation, Affinity Chromatography, Immunoprecipitation or Increased Temperature for Protein Isolation, Drug Interaction, and Proteomic and Peptidomic Evaluation -- Lysis Buffer Choices are Key Considerations to Ensure Effective Sample Solubilization for Protein Electrophoresis -- The Cydex Blue Assay: A One Step Protein Assay for Samples Prior to SDS Electrophoresis -- Cellulose-Acetate Electrophoresis of Hemoglobin -- Native Polyacrylamide Gels -- Isoelectric Focusing on Non-Denaturing Gels -- Determination of protein Molecular weights on SDS-PAGE -- Two-Dimensional Gel Electrophoresis by Glass Tube-Based IEF and SDS-PAGE -- Cationic Electrophoresis -- Two-Dimensional Gel Electrophoresis with Immobilized pH Gradients -- SARCOSYL-PAGE: Optimized Protocols for the Separation and Immunological Detection of PEGylated Proteins -- Tricine-SDS-PAGE -- Analysis of Protein Glycation using Phenylboronate Acrylamide Gel Electrophoresis -- Immunofixation Electrophoresis for Identification of Proteins and Specific Antibodies -- Electrophoretic Separation of Very Large Molecular Weight Proteins in SDS-Agarose -- Increase in Local Protein Concentration by Field-Inversion Gel Electrophoresis -- Two-Dimensional Difference Gel Electrophoresis -- Immunoelectrophoresis: A Method with Many Faces -- Tris-Acetate Polyacrylamide Gradient Gels for the Simultaneous Electrophoretic Analysis of Proteins if Very High and Low Molecular Mass -- Diagnoal Electrophoresis for Detection of Proteins Involved in Disulfide Bonds -- Identification of Proteins on Archived 2D Gels -- Two-Dimensional Gel Electrophoresis: Vertical Isoelectric Focusing -- A Foodomics Approach: CE-MS for Comparative Metabolomics of Colon Cancer Cells Treated with Dietary Polyphenols -- Characterization of New Cyclic D, L – α – Alternate Amino Acid Peptides by Capillary Electrophoresis Coupled to Electrospray Ionization Mass Spectrometry -- “Microchip Electrophoresis”, with Respect to “Profiling of Aß Peptides in the Cerebrospinal Fluid of Patients with Alzheier’s Disease” -- Measuring Low-Picomolar Apparent Binding Affinities by Minigel Electrophoretic Mobility Shift -- Identification of Proteins Interacting with Single Nucleotide Polymorphisms (SNPs) by DNA Pull-Down Assay -- Horizontal Agarose Gel Mobility Shift Assay for Protein-RNA Complexes -- Applications of Immobilized Metal affinity Electrophoresis -- Isoelectric Focusing in Agarose Gel for Detection of Oligoclonal Bands in Cerebrospinal and Other Biological Fluids -- A Combined Free Flow Electrophoresis and DIGE Approach to Compare Proteins in Complex Biological Samples -- SDS PAGE for 35S Immunoprecipitation and Immunoprecipitation Western Blotting -- A Multichannel Gel Electrophoresis and Continuous Fraction Collection Apparatus for High-Throughput Protein Separation and Characterization -- Cell Surface Protein Biotinylation for SDS-PAGE Analysis -- Isolation of Proteins from Polyacrylamide Gels -- Continuous Elution SDS-PAGE with a Modified Standard Gel Apparatus to Separate and Isolate an Array of Proteins from Complex Mixtures -- Protein Extraction from Gels: A Brief Review -- Gel Absorption-Based Sample Preparation Method for Shotgun Analysis of Membrane Proteome -- Ultra-Rapid Sodium Dodecyl Sulfate Polyacrylamide Mini-Gel Electrophoresis -- A Brief Review of Other Notable Electrophoretic Methods -- Single-Cell High Resolution Detection and Quantification of Protein Isoforms Differing by a Single Charge Unit -- Artifacts and Common Errors in Protein Gel Electrophoresis
    Abstract: This volume expands upon Protein Electrophoresis (2012) and provides readers with easy-to-follow and reproducible methods to study electrophoresis. The chapters in this book cover topics such as the Cydex Blue assay; cellulose-acetate electrophoresis of hemoglobin; cationic electrophoresis; tricine-SDS-Page; identification of proteins on archived 2-D gels; cell surface protein biotinylation of SDS-PAGE analysis; and artifacts and common errors in protein gel electrophoresis. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Practical and thorough, Electrophoretic Separation of Proteins: Methods and Protocols is a valuable resource for researchers who are interested in learning and experimenting with this field
    Pages: XV, 523 p. 112 illus., 61 illus. in color. : online resource.
    ISBN: 9781493987931
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  • 4
    Keywords: Life sciences ; Biochemistry ; Life sciences ; Protein Science ; Springer eBooks
    Description / Table of Contents: The Early Days of Blotting -- Origins of Protein Blotting -- Western Blotting: Remembrance of Things Past -- Simian Virus 40 and Protein Transfer -- Western Blotting: An Introduction -- From Little Helpers to Automation -- Spectrophotometric Methods to Determine Protein Concentration -- Solubilization of Proteins: The Importance of Lysis Buffer Choice -- Simultaneous Immunoblotting Analysis with Activity Gel Electrophoresis and 2-D Gel Electrophoresis -- Diffusion Blotting: A Rapid and Simple Method for Production of Multiple Blots from a Single Gel -- Multiple Immunoblots by Passive Diffusion of Proteins from a Single SDS-PAGE Gel -- Slice Blotting -- Localizing Proteins by Tissue Printing -- Dot-Immunobinding Assay (Dot-Iba) -- Analysis of Antibody Clonotype by Affinity Immunoblotting -- Glycosaminoglycan Blotting and Detection after Electrophoresis Separation -- A Well-Based Reverse-Phase Protein Array of Formalin-Fixed Paraffin-Embedded Tissue -- Quantitative Computerized Western Blotting in Detail -- Cationic Electrophoresis and Eastern Blotting -- A Miniaturized Blotting System for Simultaneous Detecting of Different Autoantibodies -- Proteomic Expressional Profiling of a Paraffin Embedded Tissue by Multiplex Tissue Immunoblotting -- Post-Staining Electroblotting for Efficient and Reliable Peptide Blotting -- Multistrip Western Blotting: A Tool for Comparative Quantitative Analysis of Multiple Proteins -- Western Blotting using PVDF Membranes and Its Downstream Applications -- Blotting from PhastGel to Membranes by Ultrasound -- Western Blotting of High and Low Molecular Weight Proteins Using Heat -- Membrane Strip Affinity Purification of Autoantibodies -- Strip Immunoblotting of Multiple Antigenic Peptides -- Double-Blotting: A Solution to the Problem of Non-Specific Binding of Secondary Antibodies in Immunoblotting Procedures -- Method for Resolution and Western Blotting of Very Large Proteins Using Agarose Electrophoresis -- Immunodetection of P-Selectin Using an Antibody to its C-Terminal Tag -- Improvements and Variants of the Mutliple Antigen Blot Assay-MABA: An Immunoenzymatic Technique for Simultaneous Antigen and Antibody Screening -- Blotting from Immobilized pH Gradient Gels: Application to Total Cell Lysates -- Immunoprecipitation: Western Blot for Proteins of Low Abundance -- Native Electrophoresis and Western Blot Analysis (NEWeB): Methods and Applications -- Shift-Western Blotting: Separate Analysis of Protein and DNA from Protein-DNA Complexes -- Grid-Immunoblotting -- Detection and Quantification of Protein-Protein Interactions by Far-Western Blotting -- Western Blot Analysis of Adhesive Interactions under Fluid Shear Conditions: The Blot Rolling Assay -- Centrifuge Blotting -- Blotting of Coomassie Blue Stained Proteins from PAGE Gels to Transparencies -- B Cell ELISPOT: For the Identification of Antigen-Specific Antibody Secreting Cells -- T Cell ELISPOT: For the Identification of Specific Cytokine-Secreting T Cells -- Membrane Microplates for One- and Two-Color ELISPOT and FLUOROSPOT Assays -- SDS PAGE to Immunoblot in One Hour -- Single Cell Western Blotting -- Protein Detection by Simple Westerń„Ø Analysis -- Western Blotting Using Microfluidics -- Two-Dimensional Gel-Based Protein Standardization Verified by Western Blot Analysis -- Fingerprint Deposition of Nitrocellulose and Polyvinylidene Difluoride Membranes Using Alkaline Phosphatase -- Other Notable Protein Blotting Methods: A Brief Review.℗ ℗ ℗ ℗ ℗ ℗ ℗ ℗ ℗ ℗
    Abstract: This volume covers past and present western blot techniques, such as diffusion blotting, slice blotting, blotting of high and low molecular weight proteins, single cell blotting and automated blotting. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Thorough and cutting-edge, Western Blotting: Methods and Protocols will serve as an invaluable reference for those interested in further study into this fascinating field. ℗
    Pages: XIV, 509 p. 140 illus., 65 illus. in color. : online resource.
    ISBN: 9781493926947
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  • 5
    Keywords: Life sciences ; Biochemistry ; Life sciences ; Protein Science ; Springer eBooks
    Description / Table of Contents: Western Blotting: Origin and Ascent of the Species -- Methods to Concentrate Proteins for Protein Isolation, Proteomic and Peptidomic Evaluation -- Measuring Protein Concentration on Nitrocellulose and After the Electrophoretic Transfer of Protein to Nitrocellulose -- Detection of Blotted Proteins: Not all Blockers are Created Equal -- Protein Stains to Detect Antigen on Membranes -- Fluorescent Labeling of Proteins and Its Application to SDS-PAGE and Western Blotting -- Rapid, Antibody-free Detection of Recombinant Proteins on Blots Using Enzyme Fragment Complementation -- Use of Non-Radioactive Detection Method for North- and South-Western Blot -- ℗ Immunoblotting using Radiolabeled Reagents for Detection -- Immunoblotting of Antigens: Whole, Strip and New-Line Nitrocellulose Membrane Immunoblotting Using the Chemiluminescence Technique -- Detection of Protein Carbonyls by Means of Biotin Hydrazide ́€“Streptavidin Affinity Methods -- Direct Immunodetection of Antigens within the Precast Polyacrylamide Gel -- Quantitative Analysis of Signal Transduction with In-Cell Western Immunofluorescence Assays -- Ultrasensitive Protein Detection on Dot Blots and Western Blots with Semiconducting Polymer Dots -- Co-detection of Target and Total Protein by CyDye Labeling and Fluorescent ECL-plex Immunoblotting in a Standard Proteomics Workflow -- Using Biotinylated Proteins to Demonstrate Immunodetection of Antigens via Western Blotting, Dot blots and Immunohistochemistry -- Calcium Binding by Ro 60 Multiple Antigenic Peptides on PVDF Membrane -- Sequential use of Immunoblots for Characterization of Autoantibody Specificities -- Nanogold Immunodetection Detection Systems for the Identification of Autoantigens by Western Blotting -- Application of Intermittent Microwave Irradiation to Western Blot Analysis -- Visualization of Unstained Protein Bands on PVDF -- Multiplexed Fluorescent Immunodetection Using Low Auto-Fluorescence Immobilon℗ʼ-FL Membrane -- Cold Microwave-Enabled Protein Detection and Quantification -- TLC-Blot (Far-Eastern Blot) and Its Application to Functional Lipidomics -- Analysis of Electroblotted Proteins by Mass Spectrometry -- On Membrane Renaturation of Recombinant Ro60 Autoantigen by Calcium Ions -- Phosphoprotein Detection on Protein Electroblot Using a Phosphate-Specific Fluorophore -- Purification of Tryptic Digests on Polyvinylidene Difluoride Membrane -- Detection of Blotted Proteins on Nitrocellulose/PVDF Membranes by Alta -- Non-stripping ́€œRainboẃ€ and Multiple Antigen Detection (MAD) Western Blotting. - Supported Molecular Matrix Electrophoresis -- Parafilm-M℗ʼ, an Available Cost Effective Alternative for Immuno-blot Pouches -- Succinylation-Alcian Blue Staining of Mucins on Polyvinylidene Difluoride Membranes -- Comparison of Chemiluminescence vs. Infrared Techniques for Detection of Fetuin-A in Saliva -- A Novel Methodology for Stripping and Reprobing of Western Blots Originally Developed with Colorimetric Substrate TMB -- Other Notable Methods of Membrane Protein Detection: A Brief Review -- Nitrocellulose Membrane: The New Canvas -- Invisible Ink Marking in ECL Membrane Assays. ℗ ℗
    Abstract: This detailed volume provides methods and techniques for detection after blotting.℗ Chapters guide readers through a number of variations on the theme of protein transfer to solid support followed by detection, presenting adaptations of traditional techniques, and original methods of protein blotting. Written for the Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Practical and authoritative, Detection of Blotted Proteins: Methods and Protocols presents numerous techniques based on the Western blot, providing detailed, readily reproducible methods, tips, and alternatives directly and easily transferable to the laboratory setting
    Pages: XIV, 386 p. 101 illus., 48 illus. in color. : online resource.
    ISBN: 9781493927180
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  • 6
    ISSN: 1573-2592
    Keywords: Systemic lupus erythematosus ; autoantibodies ; autoantigens ; RNA-protein ; Ro ; SS-A ; etiology ; vesicular stomatitis virus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1573-2592
    Keywords: Ro ; SS-A ; anti-Ro ; epitope ; systemic lupus erythematosus ; Sjögren's syndrome ; vesicular stomatitis virus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The Ro ribonucleoprotein is composed of hY RNA and a 60.7-kD peptide that is antigenic for autoantibodies produced by many patients with systemic lupus erythematosus or Sjögren's syndrome and mothers of newborns with complete congenital heart block. A major immunoreactive fragment (13 kD) of the 60-kD Ro is bound by 28 of 45 (62%) of the anti-Ro sera tested. Amino acid sequence analysis localizes this fragment to the carboxyl end of the 60-kD Ro peptide. All possible overlapping octapeptides of this 13-kD peptide of 60-kD Ro have been assessed for antigenicity. Sera that bind the 13-kD peptide fragment in immunoblot generally also bind the octapeptides of Ro spanning the sequence AIALREYRKKMDIPA (P<0.01). Inhibition studies with synthetic peptides and purified Ro have established specificity for reference serum antibody binding to an antigenic octapeptide, EYRKKMDI, from this region. The closely related sequence EYRKKLMD is found in the nucleocapsid protein of vesicular stomatitis virus and may portend an immunologic link to this or a related viral antigen. These results also demonstrate that despite fine specificity variation between human sera, there are recurring patterns of anti-Ro binding shared by some patients who have precipitating anti-Ro autoantibodies.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1573-2592
    Keywords: Systemic lupus erythematosus ; autoantibodies ; autoantigens ; antinuclear antibodies
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A gel filtration method was developed to estimate the number of conformational epitopes on the 60-kD Ro antigen. Anti-Ro Fab or Fab′ was incubated with native Ro antigen at different ratios and the Stokes radius molecular weight of complexes was estimated by gel filtration. Binding was saturated at 9 to 11 Fab molecules per bovine Ro molecule. Two additional Fab or Fab′ were bound if human Ro was used as the antigen. Isolated Ro antigen/anti-Ro Fab complexes were evaluated for the relative proportion of antigen to antibody at saturation of antigen with antibody and thus stoichiometry was determined. This provided data supporting there being between 7 and 11 binding sites, results similar to those with the gel filtration method. Experiments carried out with anti-Ro monoclonal antibodies showed one binding site per molecule of 60-kD Ro. Therefore, we have developed methods to count conformational epitopes on autoantigens and have applied it to the Ro/anti-Ro system. The data indicate that multiple conformational epitopes can be bound simultaneously by polyclonal anti-Ro sera from patients with systemic lupus erythematosus.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Electrophoresis 19 (1998), S. 1659-1661 
    ISSN: 0173-0835
    Keywords: Multiple antigenic peptides ; Immunoblotting ; Sodium dodecyl sulfate-polyacrylamide gel electrophoresis ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Multiple antigenic peptides (MAPs) can be efficiently separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to a nitrocellulose membrane for subsequent use in immunoblot (IgG and IgM). MAPs can be stained by Coomassie and silver on SDS-PAGE as well as by Fast Green on an immunoblot. Affinity immunoblotting for analysis of antibody clonotype distribution has also been carried out using MAPs. High performance liquid chromatography purification of the MAPs is mainly responsible for their migration as sharp bands in SDS-PAGE and immunoblotting.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
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