Labeling density

Yokota, S. (1988) Effect of particle size on labeling density for catalase in protein A-Gold immunocyto-chemistry./. Histochem. Cytochem. >6, 107-109. 

Abstract The emergence of a family of computational methods, known under the label density... 

It is worthwhile to indicate here that the label density functional theory might be used in two different contexts pragmatic and methodological. 

Constraints imposed by the intended application may limit the choice of labeling methods (Table 7.2). For example, when the probe should be intact, such as in Sj mapping, labeling by nick translation should be avoided. In contrast, for most slot blot and in situ hybridizations, nick translation or random primer extension methods provide suitable probes with an optimum label density, often with the added benefit of ss extensions on the duplex offering the possibility of amplification through hyperpolymer formation. 

Incorporation of secondary labels by random primer extension The protocol for the incorporation of secondary, nonradioactive labels differs in several respects from that for radiolabels (Table 7.16B) since (i) tagged precursor dNTPs can be bulky and less suitable as a substrate and (ii) the label density usually should be less (Section 7.6.1.2). In contrast to the protocol of radiolabeling, the concentration of each dNTP largely exceeds its although one of the dNTPs is partially replaced by labeled precursor. Consequently, it is easier to make large quantities without decreasing the specific activity. 

Song L, van Gijlswijk RP, Young IT, Tanke HJ (1997) Influence of fluorochrome labeling density... 

Fig. 14.4) were used to label oligonucleotides by Randolph et al [71]. While Cy3 contains a bis-sulfonate functionality for improved water-solubility, Cy3NOS is mono-sulfonated. The reaction of 14 and 15 produced the carboxy-substituted dye 16, which was further treated with N,N -disuccinimidyl carbonate in a mixture of DMF/pyridine to give the nonsymmetric N-hydroxysuccinimidyl substituted Cy dyes 17. Dyes 17 were then covalently linked via a short or long tether at the C-5 position of deoxyuridine to multiply-label DNA. The correlation study between the labeling density and the sensitivity of the DNA fluorescent probe demonstrated that labeling at every 6th base pair with Cy3 showed the optimal fluorescence. 

The probe/vector DNA template must be linearized to restnct the transcription to the insert. Cotranscription of the vector will reduce the label density of the probe and may cause background,... 

Chloroplast Region of lumen Total counts Total length of lumen (/ m) Labeling densities Total counts /total length of lumen (counts//im)... 

After writing a title in cell Al, enter the constants gq to g in column A. Column B is labeled Temp (°C) and column C is labeled Density (g/mL) . Enter values of temperature in column B. In cell C4, type the formula = A 5 + A 7 B4 + A 9 B4 2 + A 11 B4 3 , which uses the exponent symbol to compute and T. When you enter the formula, the number 0.999 97 is computed in cell C4. The remainder of column C is completed with a Fill Down command. The spreadsheet is not finished until it is documented by entering text in cells A13 and A14 to show what formula was used in column C. 

The measurement itself would consist of illuminating the entire sample at a donor excitation wavelength and observing the resultant acceptor fluorescence intensity. In the reasonably attainable approximations that a) the bulk densities of the two lightly-labelled polymers are the same and b) the label density (labels/volume, ) on each side of the original Interface is the same, the observed fluorescence intensity, I(t), will increase with time like ... 

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