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Additional info for Advanced Calculus of real valued functions of real variable and vectored valued functions of a vector 0
BLOTTING (i) Add transfer buffer to the blotting apparatus, which already contains the cassette with porous pads (Scotch-Brite). In this way the pads are well wetted. l). Reuse of transfer buffer may be possible (Gooderham, 1984). Take out the cassette and pads and lay them horizontally on a tray. Open up the cassette so that the mechanical support and one pad are facing up; set aside the excess pads and the second half of the cassette. If individual lanes are to be blotted separately, cut the gel accordingly.
For example, NC is dissolved in acetone or high concentrations (> 90%) of methanol. Thus a polypeptide-containing piece of NC is dissolved and the resultant slurry is either extracted to obtain the protein in an aqueous phase or simply mixed with adjuvant and used directly. Clearly a “smart membrane” would be useful. Such a membrane would bind protein via a reversible cross-linking moiety. But alas, as long as blotting matrices are in essence “misused” filtration media, we blottologists will simply have to make do.
Washes should consist of frequent changes of the wash solution. The presence of quenchers does not usually add to the wash efficiency. On the other hand, nonionic detergents usually reduce the extent of background. The minimal adequate wash should be determined. Excessive washing may lead to loss of 44 JONATHAN M. GERSHONI signal, especially when the protein-ligand complex is of low avidity. Washing with ice-cold buffer may help in this respect. (v) S e c d probe. If a second probe is necessary, it should be calibrated in the same manner as described in step iii.
Advanced Calculus of real valued functions of real variable and vectored valued functions of a vector 0