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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.

Western Blotting: Methods and Protocols

1. The Early Days of Blotting
Edwin Southern

2. Origins of Protein Blotting
Harry Towbin

3. Western Blotting: Remembrance of Things Past
W. Neal Burnette

4. Simian Virus 40 and Protein Transfer
Jaime Renart

5. Western Blotting: An Introduction
Biji T. Kurien and R. Hal Scofield

6. From Little Helpers to Automation
Harry Towbin

7. Spectrophotometric Methods to Determine Protein Concentration
J.P. Dean Goldring

8. Solubilization of Proteins: The Importance of Lysis Buffer Choice
Mandy Peach, Noelle Marsh, Ewa I. Miskiewicz, and Daniel J. MacPhee

9. Simultaneous Immunoblotting Analysis with Activity Gel Electrophoresis and 2-D Gel Electrophoresis
Der-Yen Lee and Geen-Dong Chang

10. Diffusion Blotting: A Rapid and Simple Method for Production of Multiple Blots from a Single Gel
Ingrid Olsen and Harald G. Wiker

11. Multiple Immunoblots by Passive Diffusion of Proteins from a Single SDS-PAGE Gel
Biji T. Kurien and R. Hal Scofield

12. Slice Blotting
Graeme Lowe

13. Localizing Proteins by Tissue Printing
Rafael F. Pont-Lezica

14. Dot-Immunobinding Assay (Dot-Iba)
Sumi Surendran, Annamma Mathai, and Vishnampet Venkataraman Radhakrishnan

15. Analysis of Antibody Clonotype by Affinity Immunoblotting
Biji T. Kurien and R. Hal Scofield

16. Glycosaminoglycan Blotting and Detection after Electrophoresis Separation
Nicola Volpi and Francesca Maccari

17. A Well-Based Reverse-Phase Protein Array of Formalin-Fixed Paraffin-Embedded Tissue
Joon-Yong Chung and Stephen M. Hewitt

18. Quantitative Computerized Western Blotting in Detail
Dalit Talmi-Frank, Charles L. Jaffe, and Gad Baneth

19. Cationic Electrophoresis and Eastern Blotting
Engelbert Buxbaum

20. A Miniaturized Blotting System for Simultaneous Detecting of Different Autoantibodies
Ulrich Canzler, Holger Bartsch, Kai Großmann, Werner Lehmann, Karsten Conrad, Biji T. Kurien, Yaser Dorri, R. Hal Scofield, and Michael P. Bachmann

21. Proteomic Expressional Profiling of a Paraffin Embedded Tissue by Multiplex Tissue Immunoblotting
Joon-Yong Chung and Stephen M. Hewitt

22. Post-Staining Electroblotting for Efficient and Reliable Peptide Blotting
Der-Yen Lee and Geen-Dong Chang

23. Multistrip Western Blotting: A Tool for Comparative Quantitative Analysis of Multiple Proteins
Edita Aksamitiene, Jan B. Hoek, and Anatoly Kiyatkin

24. Western Blotting using PVDF Membranes and Its Downstream Applications
Setsuko Komatsu

25. Blotting from PhastGel to Membranes by Ultrasound
Joseph Kost and Aharon Azagury

26. Western Blotting of High and Low Molecular Weight Proteins Using Heat
Biji T. Kuien and R. Hal Scofield

27. Membrane Strip Affinity Purification of Autoantibodies
Biji T. Kurien

28. Strip Immunoblotting of Multiple Antigenic Peptides
Biji T. Kurien

29. Double-Blotting: A Solution to the Problem of Non-Specific Binding of Secondary Antibodies in Immunoblotting Procedures
Françoise Lasne

30. Method for Resolution and Western Blotting of Very Large Proteins Using Agarose Electrophoresis
Marion L. Greaser and Chad M. Warren

31. Immunodetection of P-Selectin Using an Antibody to its C-Terminal Tag
Padmaja Mehta

32. Improvements and Variants of the Mutliple Antigen Blot Assay-MABA: An Immunoenzymatic Technique for Simultaneous Antigen and Antibody Screening
Oscar Noya, Sandra Losada, Marilyan Toledo, Adriana Gauna, María Angelita Lorenzo, Henry Bermúdez, and Belkisyolé Alarcón de Noya

33. Blotting from Immobilized pH Gradient Gels: Application to Total Cell Lysates
Harry Towbin

34. Immunoprecipitation: Western Blot for Proteins of Low Abundance
Edward P. Trieu and Ira N. Targoff

35. Native Electrophoresis and Western Blot Analysis (NEWeB): Methods and Applications
Ioannis N. Manoussopoulos and Mina Tsagris

36. Shift-Western Blotting: Separate Analysis of Protein and DNA from Protein-DNA Complexes
Matthias Harbers

37. Grid-Immunoblotting
Olga Yeliosof and R. Hal Scofield

38. Detection and Quantification of Protein-Protein Interactions by Far-Western Blotting
Joshua A. Jadwin, Bruce J. Mayer, and Kazuya Machida

39. Western Blot Analysis of Adhesive Interactions under Fluid Shear Conditions: The Blot Rolling Assay
Robert Sackstein and Robert Fuhlbrigge

40. Centrifuge Blotting
Jinny Paul

41. Blotting of Coomassie Blue Stained Proteins from PAGE Gels to Transparencies
Jinny Paul

42. B Cell ELISPOT: For the Identification of Antigen-Specific Antibody Secreting Cells
Hemangi B. Shah and Kristi A. Koelsch

43. T Cell ELISPOT: For the Identification of Specific Cytokine-Secreting T Cells
Kerry M. Leehan and Kristi A. Koelsch

44. Membrane Microplates for One- and Two-Color ELISPOT and FLUOROSPOT Assays
Alexander E. Kalyuzhny
45. SDS PAGE to Immunoblot in One Hour
Biji T. Kurien, Debashish Danda, Michael Bachmann, and R. Hal Scofield

46. Single Cell Western Blotting
Syed MS Quadri

47. Protein Detection by Simple Western(TM) Analysis
Valerie Harris

48. Western Blotting Using Microfluidics
Pothur R. Srinivas

49. Two-Dimensional Gel-Based Protein Standardization Verified by Western Blot Analysis
Hisao Haniu, Daisuke Watanabe, Yusuke Kawashima, and Hiroyuki Matsumoto

50. Fingerprint Deposition of Nitrocellulose and Polyvinylidene Difluoride Membranes Using Alkaline Phosphatase
Biji T. Kurien, Debashish Danda, and R. Hal Scofield

51. Other Notable Protein Blotting Methods: A Brief Review
Biji T. Kurien and R. Hal Scofield