Protein Blotting

Electrophoresis and Blotting

Protein Blotting Guide

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Protein Blotting Guide

Theory and Products

Part 1 Theory and Products
Chapter 1 Overview of Protein Blotting
Transfer Detection General Considerations and Workflow

5 5 6 6 6 9 10 10 10 11 12 12 12 12 13 13 14 14 14 14 15 15 15 17 18 18 18 18 18 19 19 19 20 20 20 21 21 23 24 24 24 24 24 24 26 26 26 26 26 26

Chapter 5Detection and Imaging
Total Protein Detection
Anionic Dyes Fluorescent Protein Stains Stain-Free Technology Colloidal Gold

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Transfer Buffer Formulations
Towbin Buffer Towbin Buffer with SDS Bjerrum Schafer-Nielsen BufferBjerrum Schafer-Nielsen Buffer with SDS CAPS Buffer Dunn Carbonate Buffer 0.7% Acetic Acid

Immunodetection
Immunodetection Workflow
Blocking Antibody Incubations Washes

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Chapter 2 Methods and Instrumentation
Protein Blotting Methods
Electrophoretic Transfer
Tank Blotting Semi-DryBlotting

Detection Buffer Formulations
General Detection Buffers Total Protein Staining Buffers and Solutions Substrate Buffers and Solutions Stripping Buffer

Antibody Selection and Dilution
Primary Antibodies Species-Specific Secondary Antibodies Antibody-Specific Ligands

Microfiltration (Dot Blotting)

Blotting Systems and Power Supplies
Tank Blotting Cells
Mini Trans-Blot® Cell andCriterion™ Blotter Trans-Blot® Cell Trans-Blot® Plus Cell

Detection Methods
Colorimetric Detection Premixed and Individual Colorimetric Substrates Immun-Blot® Assay Kits Immun-Blot Amplified AP Kit Opti-4CN™ and Amplified Opti-4CN Substrate and Detection Kits Chemiluminescence Detection Immun-Star ™ Chemiluminescence Kits Fluorescence Detection Other Detection Methods BioluminescenceChemifluorescence Autoradiography Immunogold Labeling Stripping and Reprobing

Part 3 Troubleshooting
Transfer
Electrophoretic Transfer Microfiltration

Semi-Dry Blotting Cells
Trans-Blot® SD Semi-Dry Cell Trans-Blot® Turbo™ System

Detection
Immunodetection Multiscreen Apparatus Total Protein Detection

Microfiltration Apparatus
Bio-Dot® and Bio-Dot® SF Apparatus

Power Supplies forElectrophoretic Transfers
PowerPac™ HC Power Supply PowerPac™ Universal Power Supply

Appendix
Protein Standards for Blotting
Unstained Standards for Protein Blotting
Unstained SDS-PAGE Standards Precision Plus Protein Unstained Standards

Chapter 3 Membranes and Transfer Buffers
Membranes and Blotting Papers
Nitrocellulose and Supported Nitrocellulose Polyvinylidene Difluoride (PVDF)Immun-Blot® and Immun-Blot LF PVDF for Western Blotting Sequi-Blot ™ PVDF for Protein Sequencing

Imaging — Analysis and Documentation
Luminescence Detection Digital Imaging for Fluorescence, Chemifluorescence, and Colorimetric Detection Autoradiography Analysis Software

Prestained Standards for Western Blotting
Precision Plus Protein Prestained Standards Kaleidoscope Standards Prestained SDS-PAGEStandards

Blotting Filter Papers Membrane/Filter Paper Sandwiches

Precision Plus Protein™ WesternC™ Standards

Transfer Buffers
Towbin and Bjerrum Schafer-Nielsen Buffers (Tris/Glycine Buffers) CAPS Buffer Discontinuous Tris-CAPS Buffer System (for Semi-Dry Transfer) Dunn Carbonate Buffer Other Buffers

Glossary References and Related Reading Ordering Information

Part 2 MethodsProtocols
Electrophoretic Transfers
Reagent and Materials Preparation Tank Blotting Procedure Prepare the Gel and Membrane Sandwich Assemble the Tank and Program the Power Supply Semi-Dry Blotting Procedure Trans-Blot® Turbo™ Blotting Procedure

Chapter 4 Transfer Conditions
General Workflow – Electrophoretic Transfer Power Conditions
Useful Equations Joule Heating and Other Factors Affecting...