Biology Reference
In-Depth Information
CHAPTER
12
Memb
rane Isolation M
ethods
OUTLINE
A. Introduction
239
D. Membrane Fractionation: Non-
Centrifugation Methods
Microsomes
240
253
Affinity Chromatography
253
Membrane Isolation Steps
241
Lectin-Affinity Chromatography
254
B. Breaking Open the Cell:
Homogenization
Antibody-Affinity Chromatography
256
241
Ligand-Receptor Affinity
Chromatography
Homogenization Methods
242
256
1. Use of Enzymes
242
Anion Exchange Chromatography
256
2. Shear Force (Mortar and Pestle)
242
Derivitized Beads
257
3. Blenders
242
Magnetic Beads
257
4. Osmotic Gradients
244
Other Types of Microbeads
258
5. Sonication
245
Other Methodologies
258
6. Bead Beaters
246
Two Phase Partitioning
258
7. Gas Ebullition (High Pressure
'Bomb')
Silica Particles
259
246
Separation by Size
259
8. French Press
247
Membrane-Specific 'Tricks'
259
9. Freeze/Thaw
248
E. Membrane Markers
260
C. Membrane Fractionation:
Centrifugation
248
Summary
261
Differential Centrifugation
248
References
262
Density Gradient Centrifugation
249
A. INTRODUCTION
Isolating biological membranes is as much an art form as it is a science. The procedures are
tedious and require patience, precision, and organization. Since every membrane has its own
