Preparation of Dual-Color Polarity-Marked Fluorescent Microtubule Seeds
Assaying microtubule dynamics in vitro requires stabilized nucleation centers, a method to immobilize individual microtubules onto a surface, and a specialized microscope to image the microtubule. Microtubules are polar structures with different dynamic properties at the plus and minus ends. However, the dynamics of the two ends can be modified by the addition of other proteins, such as microtubule plus-end-tracking proteins (+TIPs), so that it becomes impossible to distinguish the microtubule polarity by measuring the differences in the dynamic properties of the ends alone. In this chapter, we describe a method for labeling tubulin protein with N -hydroxysuccinimide ester fluorescent dyes, enabling the formation of dual-color polarity-marked stable microtubule seeds that can be immobilized onto a microscopic cover glass for imaging by fluorescence microscopy. These seeds create functional nucleation centers for the growth of dynamic microtubules.
- CSF Extract Prep for Spindle Assembly
- DNA Fragmentation Assays for Apoptosis
- Embryonic Stem Cells as a Model for the Physiological Analysis of the Cardiovascular System
- Production and Purification of Monoclonal Antibodies
- Prospective Isolation of Mesenchymal Stem Cells from Mouse Compact Bone
- Methods to Analyze Transglutamination of Proteins Involved in Apoptosis
- Identification of Urine Organic Acids for the Detection of Inborn Errors of Metabolism Using Urease and Gas Chromatography-Mass
- 大肠杆菌细胞超声波破碎
- 小鼠白细胞介素33(IL-33)酶联免疫分析
- 细胞爬片实验经验总结