Author: Jose-Antonio Rodriguez-Rodriguez; Kara L. McKinley; Vitali Sikirzhytski; Jennifer Corona; John Maciejowski; Alexey Khodjakov; Iain M. Cheeseman; Prasad V. Jallepalli
Title: The RZZ complex integrates spindle checkpoint maintenance with dynamic expansion of unattached kinetochores Document date: 2018_4_9
ID: 0c4cbwqq_54
Snippet: The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. . https://doi.org/10.1101/297580 doi: bioRxiv preprint 24 glycerophosphate, 1 mM DTT, 0.2 mM microcystin, and 1x protease inhibitor cocktail (Sigma)) prior to nitrogen cavitation (1250 psi, 45 min; Parr Instruments) and centrifugation at 20,000 x g for 30 minutes. LAP-Rod was immunoprecipitated with GFP antibodies coupled to protein G-Dynabeads using bis(su.....
Document: The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. . https://doi.org/10.1101/297580 doi: bioRxiv preprint 24 glycerophosphate, 1 mM DTT, 0.2 mM microcystin, and 1x protease inhibitor cocktail (Sigma)) prior to nitrogen cavitation (1250 psi, 45 min; Parr Instruments) and centrifugation at 20,000 x g for 30 minutes. LAP-Rod was immunoprecipitated with GFP antibodies coupled to protein G-Dynabeads using bis(sulfosuccinimidyl)suberate (BS3). Zw10 was immunoprecipitated without BS3 crosslinking. Extracts and immunoprecipitates were separated by SDS-PAGE and transferred to PVDF or nitrocellulose membranes. Membranes were blocked and probed with primary antibodies and secondary antibody-HRP conjugates in 5% nonfat dry milk in TBST (Tris-buffered saline + 0.05% Tween-20) before detecting signals via enhanced chemiluminescence (Western Lightning Plus, PerkinElmer). All rights reserved. No reuse allowed without permission.
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