Abstract
This chapter discusses a brain microtubule protein preparation depleted of mitochondrial and synaptosomal components. Microtubule protein is isolated by two principal approaches: ion-exchange chromatography on DEAE-Sephadex and a recycling procedure involving warm-induced assembly and cold disassembly with intervening ultracentrifugation steps. Many microtubule purification methods apply one or both of the following methods that are capable of disrupting osmotically fragile organelles: hypotonic extraction of whole brain tissue and/or electromechanical blenders. These extraction methods contribute to destruction of brain mitochondria and synaptosomes, particularly, emulsifying effects by the lipid-rich myelinated white matter in whole brains. The sucrose method for microtubule extraction and isolation minimizes contamination. This chapter outlines a new protocol that minimizes such disruption and contamination while providing excellent yields. Additional information about the molecular characterization of sucrose purified microtubule protein is also presented in the chapter.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 51-60 |
| Number of pages | 10 |
| Journal | Methods in cell biology |
| Volume | 24 |
| Issue number | C |
| DOIs | |
| State | Published - Jan 1 1982 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Cell Biology
Cite this
A Brain Microtubule Protein Preparation Depleted of Mitochondrial and Synaptosomal Components. / Karr, Timothy L.; White, Hillary D.; Coughlin, Beth A.; Purich, Daniel L.
In: Methods in cell biology, Vol. 24, No. C, 01.01.1982, p. 51-60.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - A Brain Microtubule Protein Preparation Depleted of Mitochondrial and Synaptosomal Components
AU - Karr, Timothy L.
AU - White, Hillary D.
AU - Coughlin, Beth A.
AU - Purich, Daniel L.
PY - 1982/1/1
Y1 - 1982/1/1
N2 - This chapter discusses a brain microtubule protein preparation depleted of mitochondrial and synaptosomal components. Microtubule protein is isolated by two principal approaches: ion-exchange chromatography on DEAE-Sephadex and a recycling procedure involving warm-induced assembly and cold disassembly with intervening ultracentrifugation steps. Many microtubule purification methods apply one or both of the following methods that are capable of disrupting osmotically fragile organelles: hypotonic extraction of whole brain tissue and/or electromechanical blenders. These extraction methods contribute to destruction of brain mitochondria and synaptosomes, particularly, emulsifying effects by the lipid-rich myelinated white matter in whole brains. The sucrose method for microtubule extraction and isolation minimizes contamination. This chapter outlines a new protocol that minimizes such disruption and contamination while providing excellent yields. Additional information about the molecular characterization of sucrose purified microtubule protein is also presented in the chapter.
AB - This chapter discusses a brain microtubule protein preparation depleted of mitochondrial and synaptosomal components. Microtubule protein is isolated by two principal approaches: ion-exchange chromatography on DEAE-Sephadex and a recycling procedure involving warm-induced assembly and cold disassembly with intervening ultracentrifugation steps. Many microtubule purification methods apply one or both of the following methods that are capable of disrupting osmotically fragile organelles: hypotonic extraction of whole brain tissue and/or electromechanical blenders. These extraction methods contribute to destruction of brain mitochondria and synaptosomes, particularly, emulsifying effects by the lipid-rich myelinated white matter in whole brains. The sucrose method for microtubule extraction and isolation minimizes contamination. This chapter outlines a new protocol that minimizes such disruption and contamination while providing excellent yields. Additional information about the molecular characterization of sucrose purified microtubule protein is also presented in the chapter.
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UR - http://www.scopus.com/inward/citedby.url?scp=0020001692&partnerID=8YFLogxK
U2 - 10.1016/S0091-679X(08)60647-0
DO - 10.1016/S0091-679X(08)60647-0
M3 - Article
C2 - 7098996
AN - SCOPUS:0020001692
VL - 24
SP - 51
EP - 60
JO - Methods in Cell Biology
JF - Methods in Cell Biology
SN - 0091-679X
IS - C
ER -