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Choosing Colors The Stroke and Fill Panels Stroke Color Stroke Height Stroke Style Applying and changing fills with the Fill Panel Adjusting Stroke and Color The Pencil Tool Using the Pencil Mode pop-up options Understanding line processing The Line Tool The Oval Tool The Rectangle Tool Using the (Paint) Brush Tool Using the Brush Mode option Using the Brush Size option Using the Brush Shape option Using the Brush Lock Fill option Using the Brush Pressure option The difference between the Brush Tool and the Pencil Tool The Dropper Tool The Ink Bottle Tool The Paint Bucket Tool Using the Paint Bucket Gap Size option Using the Paint Bucket Lock Fill option Using the Paint Bucket Transform Fill option (aka the Reshape Arrow cursor) The Eraser Tool Using the Eraser Shape option Using the Eraser s Faucet option Using the Erase Mode option .

The engineering of proteins to carry polyhistidine tails (N-terminal or C-terminal His tag) in frame with their primary structure amino acid sequence is achieved by means of recombinant techniques (see 3) and confers a number of advantages. The His tag often provides a very simple means of puri cation of the protein by IMAC. Also, the use of such a small af nity sequence means that antibodies may be generated against the protein of interest without the need to remove the tag. Even insoluble proteins may be puri ed by pre-solubilising the protein in guanidinium chloride or urea and then applying the unfolded protein to metal af nity column puri cation under the same conditions as above. Refolding may then be attempted post af nity column puri cation. However, there are some drawbacks in using His-tag puri cation. For instance, the presence of the His tag may have a measurable distorting effect on the structure of the protein of interest and even on the function of the protein. Sometimes the protein of interest with the His tag is less soluble than the wild-type protein. Furthermore, the IMAC columns can leak heavy metal ions together with the protein of interest, which can lead to amino-acid side-chain damage from oxidation. This problem can be made worse by the general requirement to avoid using reducing agents such as DTT or high concentrations of ME during IMAC puri cation. Just occasionally, Co2+ has been immobilised in place of Ni2+ , via four chelating bonds, in order to increase the speci city of His-tag binding to metal ions and reduce leakage of heavy metal ions into the eluant. This system can even harness natural histidine clusters for example in the puri cation of chicken lactate dehydrogenase.

The examples in this section demonstrate a variation on the previous concept. Figure 8-4 shows a chart that displays vertical bands. Again, it s a combination chart this time an XY series combined with a column chart series.

Dissimilatory Dissimilatory Dissimilatory Dissimilatory Dissimilatory Assimilatory Assimilatory Assimilatory Dissimilatory

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Color Plate 8: This is perhaps the trickiest comparison to analyze. We had this problem when building our Web site for the Flash 4 Bible. When the animation resolved and the book was displayed at a slight, 14-degree angle, it was distorted and it was

corresponding to the well-known result that two spin-; objects may be vector coupled to give a spin-0 singlet and spin-1 triplet. This is a specific example of tensor methods in group theory; we will see below how to use such methods to determine the irreducible representations contained in the direct product of arbitrary representations of unitary groups.

Sampling and quality Distinguishing sample rate from bit resolution Calculating audio file sizes Working with audio formats