Western Blotting for the Non-Expert

This book fills the need for a simplified text covering western blotting protocols aimed not just at high school and college students, but the researcher with little to no experience in these techniques. It provides the principles, basic methodology, and tips and tricks to avoiding the common pitfalls of western blotting. The book also introduces simple protocols that can transform western blotting into a fun method, such as sending secret messages on membranes or using nitrocellulose membrane as a canvas for art.

In addition to the techniques, this book also covers the history of western blotting, which originated from the development of the blotting of DNA. It then delves into the importance of protein blotting, brought to the fore by the fact that the procedure has been evolving constantly since its inception in 1979, and the fact that the scientific community is faced with a multitude of ways and means of transferring proteins to membranes..

Provides simplified versions of 55 different Western Blotting protocols Aimed at the non-expert user Offers tips and tricks to avoiding the common pitfalls of western blotting

Auteur

Dr. Kurien completed his M.Phil. and Ph.D. studies in 1989 at the University of Madras, India, under Professor R. Selvam's mentorship. After moving from India to the United States, he first joined Dr. Hiroyuki Matsumoto's laboratory in 1989. He then joined Dr. Robert H Broyles' laboratory in 1992, both in Oklahoma City. From 1993 to 2010, he worked as an associate research scientist/senior research scientist at the Oklahoma Medical Research Foundation, Oklahoma City, with Dr. Hal Scofield and from 2010 to 2017 as associate professor of research at the University of Oklahoma Health Sciences. Currently, he works as a Professor of Research. His research interests include the study of free radical-mediated damage in systemic lupus erythematosus and Sjögren's syndrome and the role of the nutraceutical curcumin in autoimmune diseases. His publication record includes numerous publications in national and international peer-reviewed journals. Besides, he edited several volumes in Protein Blotting and Detection and Protein Electrophoresis as part of the Methods in Molecular Biology series.

Contenu

1. Western blotting: How it began.- 2. Basics of Western Blotting.- 3.Basic Calculations in Western Blotting.- 4.Choosing The Right Antibody For Western Blotting.- 5. Sample Preparation.- 6. Blocking membranes at places that do not have transferred proteins.- 7. A Basic Method for Western Blotting.- 8. Trouble Shooting Western Blotting.- 9. Using Biotin to demonstrate western and dot blotting in University Practical Classes- A Guide To Instructors.- 10. Nitrocellulose Membrane Art.- 11. The Use Of A Harmless Invisible Ink To Send Secret Messages On Nitrocellulose Membranes And As A Substitute For The Harmful Radioactive Ink Used As A Marker In ECL Membrane Assays.- 12. Fingerprinting On Nitrocellulose And Polyvinylidene Difluoride Membrane.- 13. Spectrometric Determination Of Protein For Electrophoresis.- 14. Concentrating Proteins For Western Blotting.- 15. Dissolving Proteins Using Lysis Buffer.- 16. Blotting AndDetecting Glycosaminoglycans.- 17. Passive Blotting From Tissue Slices.- 18. Localizing Proteins by Tissue Printing.- 19. Ultrasound Blotting.- 20. Obtaining Several Blots From A Single SDS-PAGE Gel By Passive Transfer.- 21. Diffusion Blotting From SDS-PAGE Gels Supported By Solid Plastic.- 22. Dot-Immunobinding.- 23. Simultaneous Immunoblotting Analysis With Activity Gel Electrophoresis.- 24. Protein Blotting By Centrifugation.- 25. Blotting And Immunostaining Of Multiple Antigenic Peptides.- 26. High Molecular Weight Protein Blotting Using Agarose Gel Electrophoresis.- 27. Cetyltrimethylammonium bromide PAGE And Eastern Blotting.- 28. Purification Of Autoantibodies Bound To An Autoantigen Immobilized On A Membrane Strip.- 29. Miniaturized Blotting.- 30. Affinity Immunoblotting.- 31. Transferring Coomassie Blue Stained Proteins from Polyacrylamide Gels To Transparencies.- 32. Protein Transfer From Immobilized pH Gradient Gels.- 33. SDS PAGE To Immunoblotting In Sixty Minutes.- 34. Western Blot Analysis Of Protein-DNA Complexes Formed DuringGel Shift Experiments.- 35. Electrophoretic Transfer Of High And Low Molecular Weight Proteins To Membranes Using Heat.- 36. Grid-immunoblotting.- 37. Far-western Blotting.- 38. Native Electrophoresis And Western Blot Analysis.- 39. Efficient Electroblotting Of Low Molecular Weight Protein After Staining With Coomassie.- 40. Avoiding Non-specific Binding Of Secondary Antibodies In Immunoblotting By Double-blotting.- 41.A Brief Introduction To Other Protein Blotting Methods.- 42. Quantitative Computerized Western Blotting In Detail.- 43. Common Problems In Electrophoresis.