Abstract
Gel‐filtration is commonly used to remove contaminants from conventional actin prepared by the method of Spudich and Watt. It has been shown that this procedure removes the majority of a factor that reduces the low‐shear viscosity of actin. We have previously reported that this factor is Cap Z, a barbed end capping protein. We now establish that, even after gel‐filtration, enough Cap Z can be present in conventionally prepared actin to affect events occurring at the barbed ends of actin filaments. We also demonstrate that the concentration of Cap Z can be reduced to more than a log below the KD for binding of Cap Z to actin by either (1) immunoabsorbtion of conventionally prepared actin with anti‐Cap Z antibodies, or (2) an additional cycle of polymerization/depolymerization followed by repeat gel‐filtration. © 1995 Wiley‐Liss, Inc.
Original language | English (US) |
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Pages (from-to) | 164-170 |
Number of pages | 7 |
Journal | Cell Motility and the Cytoskeleton |
Volume | 30 |
Issue number | 2 |
DOIs | |
State | Published - 1995 |
Keywords
- Cap Z
- actin
- antibodies
- blotting
- chickens
- immunoabsorbance
- immuno‐affinity purification
- kinetics
- methods
- muscle proteins
- purification
ASJC Scopus subject areas
- Structural Biology
- Cell Biology