Detection of Blotted Proteins

1. Front Matter

   Pages i-xiv

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2. Western Blotting: Origin and Ascent of the Species

   • W. Neal Burnette

   Pages 1-3

3. Methods to Concentrate Proteins for Protein Isolation, Proteomic, and Peptidomic Evaluation

   • J. P. Dean Goldring

   Pages 5-18

4. Measuring Protein Concentration on Nitrocellulose and After the Electrophoretic Transfer of Protein to Nitrocellulose

   • J. P. Dean Goldring

   Pages 19-25

5. Detection of Blotted Proteins: Not All Blockers Are Created Equal

   • Vishal Kothari, Suresh T. Mathews

   Pages 27-32

6. Protein Stains to Detect Antigen on Membranes

   • Anil Dsouza, R. Hal Scofield

   Pages 33-40

7. Fluorescent Labeling of Proteins and Its Application to SDS-PAGE and Western Blotting

   • F. Javier Alba, Salvador Bartolomé, Antonio Bermúdez, Joan-Ramon Daban

   Pages 41-50

8. Rapid, Antibody-Free Detection of Recombinant Proteins on Blots Using Enzyme Fragment Complementation

   • Neil W. Charter, Joe Horecka, Chin-Yee Loh, Albert Doan, Tom Wehrman, Keith R. Olson

   Pages 51-61

9. Use of Nonradioactive Detection Method for North- and South-Western Blot

   • Claudia Franke, Daniel Gräfe, Holger Bartsch, Michael P. Bachmann

   Pages 63-71

10. Immunoblotting Using Radiolabeled Reagents for Detection

    • Holger Bartsch, Claudia Franke, Michael P. Bachmann

    Pages 73-78

11. Immunoblotting of Antigens: Whole, Strip, and New-Line Nitrocellulose Membrane Immunoblotting Using the Chemiluminescence Technique

    • Yaser Dorri

    Pages 79-94

12. Detection of Protein Carbonyls by Means of Biotin Hydrazide–Streptavidin Affinity Methods

    • Kenneth Hensley

    Pages 95-100

13. Direct Immunodetection of Antigens Within the Precast Polyacrylamide Gel

    • Surbhi Desai, Boguslawa R. Dworecki, Marie C. Nlend

    Pages 101-114

14. Quantitative Analysis of Signal Transduction with In-Cell Western Immunofluorescence Assays

    • Vince Boveia, Amy Schutz-Geschwender

    Pages 115-130

15. Ultrasensitive Protein Detection on Dot Blots and Western Blots with Semiconducting Polymer Dots

    • Fangmao Ye, Polina B. Smith, Daniel T. Chiu

    Pages 131-137

16. Co-detection of Target and Total Protein by CyDye Labeling and Fluorescent ECL Plex Immunoblotting in a Standard Proteomics Workflow

    • Caitriona Scaife, Ciara A. McManus, Pamela M. Donoghue, Michael J. Dunn

    Pages 139-149

17. Using Biotinylated Proteins to Demonstrate Immunodetection of Antigens via Western Blotting, Dot Blots, and Immunohistochemistry

    • Thomas Millar, Ronald Knighton, Jo-Anne Chuck

    Pages 151-164

18. Calcium Binding by Ro 60 Multiple Antigenic Peptides on PVDF Membrane

    • Biji T. Kurien, Michael P. Bachmann

    Pages 165-171

19. Sequential Use of Immunoblots for Characterization of Autoantibody Specificities

    • Holger Bartsch, Michael P. Bachmann

    Pages 173-178

20. Nanogold Immunodetection Detection Systems for the Identification of Autoantigens by Western Blotting

    • Jacen S. Moore, R. Hal Scofield

    Pages 179-184

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.

• detecting blotted molecular species
• detecting the blotted proteins
• protein blotting
• protein transfer
• western blotting techniques

• Arthritis and Clinical Immunology Program, Oklahoma Medical Research Foundation, Oklahoma City, USA

  Biji T. Kurien

• Department of Medicine, Oklahoma Medical Research Foundation, Oklahoma City, USA

  R. Hal Scofield

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