Book chapter
[13] Purification of novel kinesins from embryonic systems
Methods in Enzymology, pp.133-154
Elsevier Science & Technology
1998
Handle:
https://hdl.handle.net/2376/104823
PMID: 9751878
Abstract
Several kinesin holoenzymes, including the heterotrimeric kinesin-II and bipolar KLP61F complexes described here, are being purified in our laboratory using microtubule affinity precipitation and conventional biochemical fractionation procedures. These protocols have been optimized by using pan-kinesin peptide antibodies and subunit-specific antibodies to monitor the enrichment of kinesin-related polypeptides in particular fractions by immunoblotting. Protein purification represents the most direct route available for determining the oligomeric state and subunit composition of a kinesin holoenzyme, for identifying tightly associated accessory subunits such as SpKAP115, and for determining the molecular architecture and functional properties of native kinesin motors. Protein purification methods therefore represent an important complementary approach to molecular genetic approaches that are being pursued in many other laboratories.
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Details
- Title
- [13] Purification of novel kinesins from embryonic systems
- Creators
- David MeyerDaniel R RinesAnna KashinaDouglas G ColeJonathan M Scholey
- Publication Details
- Methods in Enzymology, pp.133-154
- Academic Unit
- Center for Reproductive Biology
- Publisher
- Elsevier Science & Technology
- Identifiers
- 99900546842801842
- Language
- English
- Resource Type
- Book chapter