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  • 1
    Online Resource
    Online Resource
    Amsterdam : Elsevier / Academic Press
    UID:
    b3kat_BV041415350
    Format: 1 Online-Ressource
    Edition: 1. ed.
    ISBN: 9780124200678 , 9780124200944
    Series Statement: Methods in enzymology 533
    Language: English
    Subjects: Biology
    RVK:
    Keywords: Enzymologie ; Labortechnik ; Zelle ; Labortechnik ; Aufsatzsammlung ; Aufsatzsammlung
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  • 2
    Online Resource
    Online Resource
    Amsterdam : Elsevier
    UID:
    b3kat_BV041268608
    Format: 1 Online-Ressource
    ISBN: 9780124200371 , 9780124200517
    Series Statement: Methods in enzymology 530
    Language: English
    Subjects: Biology
    RVK:
    Keywords: Enzymologie ; Labortechnik ; RNS ; Aufsatzsammlung
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  • 3
    Online Resource
    Online Resource
    Amsterdam : Elsevier
    UID:
    b3kat_BV041267932
    Format: 1 Online-Ressource
    ISBN: 9780124186873 , 9780124199545
    Series Statement: Methods in enzymology 529
    Language: English
    Subjects: Biology
    RVK:
    Keywords: Enzymologie ; Labortechnik ; DNS ; Aufsatzsammlung ; Electronic books
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  • 4
    Online Resource
    Online Resource
    Amsterdam [u.a.] : Elsevier, Acad. Press
    UID:
    b3kat_BV023119555
    Format: 1 Online-Ressource
    ISBN: 9780123739698
    Series Statement: Methods in enzymology 430
    Language: English
    Subjects: Biology
    RVK:
    Keywords: Translation ; Startreaktion ; Biophysik ; Labortechnik ; Aufsatzsammlung ; Aufsatzsammlung
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  • 5
    Online Resource
    Online Resource
    Amsterdam [u.a.] : Elsevier, Acad. Press
    UID:
    b3kat_BV023119537
    Format: 1 Online-Ressource
    ISBN: 9780123739643
    Series Statement: Methods in enzymology 431
    Language: English
    Subjects: Biology
    RVK:
    Keywords: Translation ; Startreaktion ; Cytologie ; Labortechnik ; Aufsatzsammlung ; Aufsatzsammlung
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  • 6
    Online Resource
    Online Resource
    Amsterdam [u.a.] : Elsevier, Acad. Press
    UID:
    b3kat_BV023119625
    Format: 1 Online-Ressource
    ISBN: 9780123741912
    Series Statement: Methods in enzymology 429
    Language: English
    Subjects: Biology
    RVK:
    Keywords: Translation ; Startreaktion ; Molekulargenetik ; Labortechnik ; Aufsatzsammlung ; Aufsatzsammlung
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  • 7
    Online Resource
    Online Resource
    Amsterdam, [Netherlands] :Academic Press,
    UID:
    almahu_9947368846302882
    Format: 1 online resource (161 p.)
    Edition: First edition.
    ISBN: 0-12-800335-9 , 0-12-800279-4
    Series Statement: Methods in Enzymology, Volume 559
    Content: The critically acclaimed laboratory standard for almost 50 years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Each volume is eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with over 520 volumes and 40,000 chapters in the collection, much of the material is still relevant today and is truly an essential publication for researchers in all fields of life sciences, including microbiology, biochemistry, cancer research, and genetics, just to name a few. In this volume, number 545, we have brought
    Note: Description based upon print version of record. , Front Cover; Laboratory Methods in Enzymology: Protein Part D; Copyright; Contents; Contributors; Preface; Chapter One: Purification of His-Tagged Proteins; 1. Theory; 2. Equipment; 3. Materials; 3.1. Solutions & buffers; 4. Protocol; 4.1. Preparation; 4.2. Duration; 5. Protocol A: Purification of His-tagged Proteins Under Native Conditions; 5.1. Tip; 6. Step 1A Preparation of a Cleared E. coli Lysate Under Native Conditions; 6.1. Overview; 6.2. Duration; 6.3. Tip; 6.4. Tip; 6.5. Tip; 6.6. Tip; 6.7. Tip; 7. Step 2A Batch Purification of His-tagged Proteins from E. coli Under Native Conditions , 7.1. Overview7.2. Duration; 7.3. Tip; 7.4. Tip; 7.5. Tip; 7.6. Tip; 8. Step 1B Preparation of a Cleared E. coli Lysate Under Denaturing Conditions; 8.1. Overview; 8.2. Duration; 8.3. Tip; 8.4. Tip; 8.5. Tip; 8.6. Tip; 8.7. Tip; 9. Step 2B Batch Purification of His-tagged Proteins from E. coli Under Denaturing Conditions; 9.1. Overview; 9.2. Duration; 9.3. Tip; 9.4. Tip; 9.5. Tip; 9.6. Tip; References; Referenced Literature; Source References; Referenced Protocols in Methods Navigator , Chapter Two: Affinity Purification of a Recombinant Protein Expressed as a Fusion with the Maltose-Binding Protein (MBP) Tag1. Theory; 2. Equipment; 3. Materials; 3.1. Solutions & buffers; 4. Protocol; 4.1. Duration; 4.2. Preparation; 5. Step 1 Equilibration of the Column; 5.1. Overview; 5.2. Duration; 5.3. Tip; 5.4. Tip; 6. Step 2 Binding of the Protein Sample; 6.1. Overview; 6.2. Duration; 6.3. Tip; 6.4. Tip; 7. Step 3 Removal of Unbound Proteins; 7.1. Overview; 7.2. Duration; 8. Step 4 Elution of the Bound Protein; 8.1. Overview; 8.2. Duration; 8.3. Tip; 8.4. Tip; 8.5. Tip; 8.6. Tip , 8.7. Tip8.8. Tip; References; Referenced Literature; Referenced Protocols in Methods Navigator; Chapter Three: Immunoaffinity Purification of Proteins; 1. Theory; 2. Equipment; 3. Materials; 3.1. Solutions & buffers; 4. Protocol; 4.1. Duration; 4.2. Preparation; 4.3. Tip; 4.4. Tip; 5. Step 1 Equilibrate Column; 5.1. Overview; 5.2. Duration; 5.3. Tip; 5.4. Tip; 6. Step 2 Loading the Column; 6.1. Overview; 6.2. Duration; 6.3. Tip; 7. Step 3 Washing the Column; 7.1. Overview; 7.2. Duration; 7.3. Tip; 8. Step 4 Elution of the Protein from the Column; 8.1. Overview; 8.2. Duration; 8.3. Tip , 8.4. Tip8.5. Tip; 8.6. Tip; 9. Step 5 Column Regeneration and Storage; 9.1. Overview; 9.2. Duration; 9.3. Tip; References; Referenced Protocols in Methods Navigator; Chapter Four: Affinity Purification of Protein Complexes Using TAP Tags; 1. Theory; 2. Equipment; 3. Materials; 3.1. Solutions & buffers; 4. Protocol; 4.1. Preparation; 4.2. Duration; 5. Step 1 Preparation of Protein Lysates; 5.1. Overview; 5.2. Duration; 5.3. Tip; 5.4. Tip; 5.5. Tip; 6. Step 2 Binding of the Protein A Tag to IgG-Sepharose; 6.1. Overview; 6.2. Duration; 6.3. Tip; 6.4. Tip; 6.5. Tip; 6.6. Tip , 7. Step 3 TEV Protease Cleavage
    Language: English
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  • 8
    Online Resource
    Online Resource
    San Diego, California :Academic Press,
    UID:
    almahu_9947366983802882
    Format: 1 online resource (240 p.)
    ISBN: 0-12-420179-2
    Content: Laboratory Methods in Enzymology: Protein Part B brings together a number of core protocols concentrating on protein, carefully written and edited by experts.Indispensable tool for the researcher Carefully written and edited by experts to contain step-by-step protocolsIn this volume we have brought together a number of core protocols concentrating on protein
    Note: Description based upon print version of record. , Front Cover; Laboratory Methods in Enzymology: Protein Part B; Copyright; Contents; Contributors; Miscellaneous; Preface; Section I: Protein Protocols/Protein In Vitro Translation; Chapter One: In Vitro Synthesis of Proteins in Bacterial Extracts; 1. Theory; 2. Equipment; 3. Materials; 3.1. Solutions & buffers; 4. Protocol; 4.1. Duration; 4.2. Preparation; 4.3. Caution; 5. Step 1 Grow and harvest E. coli for the S30 extract; 5.1. Overview; 5.2. Duration; 5.3. Tip; 6. Step 2 Preparation of the S30 extract; 6.1. Overview; 6.2. Duration; 6.3. Tip; 6.4. Caution; 6.5. Caution , 7. Step 3 Optimization of the coupled transcription and translation reaction7.1. Overview; 7.2. Duration; 7.3. Tip; 7.4. Tip; References; Referenced Literature; Related Literature; Referenced Protocols in Methods Navigator; Chapter Two: Preparation of a Saccharomyces cerevisiae Cell-Free Extract for In Vitro Translation; 1. Theory; 2. Equipment; 3. Materials; 3.1. Solutions & buffers; 4. Protocol; 4.1. Duration; 4.2. Preparation; 5. Step 1 Preparation of Yeast Cell-Free Extract; 5.1. Overview; 5.2. Duration; 5.3. Tip; 5.4. Tip; 6. Step 2 Cell-Free Translation; 6.1. Overview; 6.2. Caution , 6.3. Duration6.4. Tip; 6.5. Tip; 6.6. Tip; 6.7. Tip; 6.8. Tip; Source References; Related Literature; Referenced Protocols in Methods Navigator; Section II: Protein Protocols/Protein In Vivo Binding Assays; Chapter Three: Yeast Two-Hybrid Screen; 1. Theory; 2. Equipment; 3. Materials; 3.1. Solutions & Buffers; 4. Protocol; 4.1. Preparation; 4.2. Construction of the DB fusion protein; 4.3. Construction of the AD fusion protein; 4.4. Testing for Self-activation of pDBLeu-X (and pPC86-Y); 4.5. Duration; 5. Step 1 Small-Scale Transformation of Yeast with pDBLeu-X; 5.1. Overview; 5.2. Duration , 6. Step 2 Two-Hybrid Screen6.1. Overview; 6.2. Duration; 7. Step 3 Confirmation of Positive Interactors; 7.1. Overview; 7.2. Duration; 8. Step 4 Plasmid Rescue from Yeast; 8.1. Overview; 8.2. Duration; 9. Step 5 Electroporation of E. coli with Yeast DNA and Identification of Positive Interactors; 9.1. Overview; 9.2. Duration; 10. Step 6 Back-Transformation of Yeast and Further Confirmation of Interactions; 10.1. Overview; 10.2. Duration; References; Related Literature; Referenced Protocols in Methods Navigator; Chapter Four: UV Cross-Linking of Interacting RNA and Protein in Cultured Cells , 1. Theory2. Equipment; 3. Materials; 3.1. Solutions & buffers; 4. Protocol; 4.1. Duration; 4.2. Preparation; 4.3. Tip; 5. Step 1 UV Cross-Link RNA-Protein Complexes; 5.1. Overview; 5.2. Duration; 5.3. Tip; 5.4. Tip; 5.5. Tip; 6. Step 2 SDS Lysis of Cells; 6.1. Overview; 6.2. Duration; 6.3. Tip; 6.4. Tip; 6.5. Tip; 7. Step 3 Immunoprecipitation; 7.1. Overview; 7.2. Duration; 7.3. Preparation; 7.4. Tip; 7.5. Tip; 8. Step 4 Proteinase K Treatment of RNA Samples; 8.1. Overview; 8.2. Duration; 8.3. Tip; 9. Step 5 RNA Analysis; 9.1. Overview; 9.2. Duration; 9.3. Tip; 10. Step 6 Protein Analysis , 10.1. Overview , English
    Additional Edition: ISBN 0-12-420120-2
    Language: English
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  • 9
    UID:
    almahu_9947366986602882
    Format: 1 online resource (lvii, 413 pages) : , illustrations (some color).
    Edition: First edition.
    ISBN: 0-12-420051-6
    Series Statement: Methods in enzymology, volume five hundred and thirty
    Content: Methods in Enzymology volumes provide an indispensable tool for the researcher. Each volume is carefully written and edited by experts to contain state-of-the-art reviews and step-by-step protocols. In this volume, we have brought together a number of core protocols concentrating on RNA, complementing the traditional content that is found in past, present and future Methods in Enzymology volumes. Indispensable tool for the researcher Carefully written and edited by experts to contain step-by-step protocolsIn this volume we have brou
    Note: "ISSN: 0076-6879." , section I. RNA protocols -- section II. In vitro transcription -- section III. Measurement of RNA synthesis and decay rates -- section IV. Polysome analysis -- section V. Reverse transcription -- section VI. RNA labeling -- section VII. RNA purification -- section VIII. RNA sequencing -- section IX. RNA structure experimental analysis. , English
    Additional Edition: ISBN 0-12-420037-0
    Additional Edition: ISBN 1-299-87154-2
    Language: English
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  • 10
    Online Resource
    Online Resource
    Amsterdam ; : Elsevier,
    UID:
    almahu_9947366711502882
    Format: 1 online resource (510 p.)
    ISBN: 1-281-04968-9 , 9786611049683 , 0-08-055318-4
    Series Statement: Methods in enzymology, v. 430
    Content: For over fifty years the Methods in Enzymology series has been the critically aclaimed laboratory standard and one of the most respected publications in the field of biochemistry. The highly relevant material makes it an essential publication for researchers in all fields of life and related sciences. This volume, the second of three on the topic of Translation Initiation includes articles written by leaders in the field.
    Note: Description based upon print version of record. , Front Cover; Translation Initiation: Reconstituted Systems and Biophysical Methods; Copyright Page; Contents; Contributors; Preface; Volume in Series; Chapter 1: Transient Kinetics, Fluorescence, and FRET in Studies of Initiation of Translation in Bacteria; 1. Introduction; 2. Experimental Outline; 3. Materials; 3.1. Stock solutions; 3.2. Reagents; 4. Experimental Procedures; 4.1. Preparation and fluorescence labeling of ribosomes and ribosomal subunits; 4.2. Zonal centrifugation; 4.3. Preparation and fluorescence labeling of initiator fMet-tRNAfMet , 4.4. Preparation of fluorescence-labeled mRNA4.5. Initiation factors; 5. Rapid Kinetic Measurements; 5.1. Measuring fluorescence intensities and FRET changes in stopped flow; 6. Quench-Flow Measurements; 7. GTPase Activity; 8. Dipeptide Formation; 9. Applications of the Method; 10. Binding of fMet-tRNA to the 30S Subunit; 11. FRET to IF3; 12. Nucleotide Binding to IF2; 13. Subunit Joining; Acknowledgments; References; Chapter 2: Binding of mRNA to the Bacterial Translation Initiation Complex; 1. Introduction; 2. Experimental Procedures; 2.1. Design of model mRNAs , 2.2. Synthesis of model mRNAs2.3. Deprotection; 2.4. Thermal melting analysis; 2.5. Labeling and purification of mRNA; 2.6. Labeling schemes; 2.7. Fluorophores; 2.8. Isolation of the 30S subunit and protein purification; 2.9. Purification of aminoacylated initiator tRNA; 2.10. Initiation complex assembly; 2.11. mRNA dissociation; 2.12. Steady-state fluorescence measurements; 2.13. Kinetic association and dissociation experiments; 2.14. Determination of equilibrium binding constants; Acknowledgments; References , Chapter 3: Real-Time Dynamics of Ribosome-Ligand Interaction by Time-Resolved Chemical Probing Methods1. Introduction; 2. General Strategy; 2.1. Buffers and mixtures; 2.2. Analysis of probing reactions; 3. Time-Resolved Chemical Probing with DMS; 3.1. Validation of time-resolved chemical probing with DMS; 3.2. Procedure for chemical probing with DMS; 3.3. Example of time-resolved chemical probing with DMS; 4. Time-Resolved Probing with ONOOK; 4.1. Validation of time-resolved probing with ONOOK; 4.2. Preparation of ONOOK; 4.3. Procedure for time-resolved probing with ONOOK , 4.4. Example of time-resolved probing with ONOOK5. Time-Resolved Chemical Probing with Fe(II)-EDTA; 5.1. Validation of time-resolved probing with Fe(II)-EDTA; 5.2. Procedure for time-resolved. probing with Fe(II)-EDTA; 5.3. Example of time-resolved probing with Fe(II)-EDTA; Acknowledgments; References; Chapter 4: Overexpression and Purification of Mammalian Mitochondrial Translational Initiation Factor 2 and Initiation Factor 3; 1. Introduction; 2. Materials; 2.1. Growth media; 2.2. Common buffers; 2.3. Buffers for IF2mt; 2.4. Buffers for IF3mt; 2.5. Reagents; 2.6. Preparation of reagents , 2.7. The high-performance liquid chromatography (HPLC) system , English
    Additional Edition: ISBN 0-12-373969-1
    Language: English
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