By Jonathan M. Scholey (Eds.)
"Motility Assays for Motor Proteins" describes handy and quantitative equipment for learning purified motor proteins similar to myosins, dyneins and kinesins. The publication additionally covers recommendations wanted for learning organelle delivery and mitotic hobbies in crude telephone extracts. every one bankruptcy provides a specific kind of motility assay in adequate aspect to permit investigators to accomplish such experiments of their personal laboratories.
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Sample text
The main limitation in this type of experiment is that a relatively weak trap has to be used to obtain a satisfactory signal-to-noise ratio, where most of the noise arises from the Brownian motion of the bead. With a weak trap, the stiffness of the trap is on the order of a few piconewtons per micrometer, which allows the bead to undergo considerable myosin-directed movement during force fluctuations. The stiffness of the trap can be enhanced considerably by the use of feedback from the bead position detector to deflect the laser beam.
6. The temperature of the assay can be increased. , 1992). 7. Other conditions for the assay that result in faster movement may be employed. For instance, tropomyosin bound to the actin filaments increases the rate of movement of human platelet myosin I1 by about twofold (see Table I). , 1990). , 1990). 2. Fast Myosins When fast-moving actin filaments are imaged by a SIT camera or a microchannel plate-intensified Newvicon camera the image often exhibits considerable lag, giving the appearance of “comet tails” trailing the bright actin filament.
064 pm. This is probably not the optimal wavelength. Berns et al. O pm), that at longer wavelengths the trap force per watt of laser power is greater and there is less damage to biological tissues. For small forces, single-mode diode lasers are an attractive and inexpensive option (Afzal and Treacy, 1992); however, NdYAG lasers, particularly diode-pumped lasers, are extremely stable. In addition, the quality of the beam is very good and they require very little maintenance or alignment. Some of them have a low-intensity noise (<1%), which means that force in the trap can be controlled to this level, though the noise is much less than the equivalent Brownian noise from a bead, so it is not a primary factor.