Antibody, double

Protocol 25.6 TUNEL-Antibody Double-labeling Method for Drosophila Embryos, 464 LASER ABLATION, 466... 

TUNEL-Antibody Double-labeling Method for Drosophila Embryos... 

Nielsen J. Haeberli G Hymenoptera venom allergy analysis of double positivity to honey bee and Vespula venom by estimation of IgE antibodies to 38 species-specific major allergens Api ml and Ves v5. Allergy 2009 64 543-548. 

The ultimate goal of microarray-based expression analysis is to acquire a comprehension of the entire cellular process, in order to exploit and to standardize the multidi-menisional relations between genotype and phenotype. However, an increasingly important parameter, which has not yet been substantially taken into account, is the role of cellular translation. This means that mRNA expression data need to be correlated with the assortment of proteins actually present in the cell. One approach is based on the use of microarrays containing double-stranded DNA probes for the analysis of DNA-protein interaction and, thus, the detection and identification of DNA-binding proteins by means of fluorescence [130] or mass spectrometry analysis [131]. Moreover, substantial efforts are currently under way to develop protein, antibody, or even cell arrays, applicable to the cor-... 

Four types of techniques for separating the bound fraction P Q from the reagent mixture are in common usage, loosely termed double antibody, solid phase, charcoal adsorption and solution precipitation. The first type is used with radioimmunoassay methods specifically, while the other three types can be used with both radioassay and radioimmunoassay methods. 

In the double antibody method of separation, a second antibody, produced by injecting the first antibody into another animal, is utilized. This antibody is used to combine with and form an insoluble complex with the first antibody. After... 

CV measurements showed that the reversible eleetrode reaetion of the [Fe(CN)6]" redox eouple was suppressed to some extent by the treatment with the DNA. The addition of the anti-DNA antibody further suppressed the redox reaetion thus decreasing the magnitudes of the CV peak currents. This is most likely caused by a steric hindrance of the bulky protein, which binds to the DNA double strands on the electrode surface, to mainly reduce the effective area of the electrode. The electrostatic repulsive effect may also contribute to the electrode response, since the isoelectric point of mouse IgM is commonly in the range of 4.5 to 7.0. Figure 11 shows the relationship between the decrease in the anodic peak current (A/p ) and the antibody concentration. As seen in this figure, the electrode system responded to the anti-DNA antibody in the concentration range of 1 — 100 nM. For the case of the mouse IgM, which does not interact with double-stranded DNA, the present system gave almost no response. The sensor did not respond to other serum proteins as well (data not shown). 

Valdivieso-Garcia, A. Riche, E. Abubakar, O. Waddell, T. E. Brooks, B. W. A double antibody sandwich enzyme-linked immunosorbent assay for the detection of Salmonella using biotinylated monoclonal antibodies. J. Food Prot. 2001, 64,1166-1171. 

Pons, J. Rajpal, A. Kirsch, J.F., Energetic analysis of an antibody/antigen interface alanine scanning mutagenesis and double mutant cycles on the HyHEL-10/lysozyme interaction, Prot. Sci. 1999, 8, 958-968. 

Ino H. Application of antigen retrieval by heating for double-label fluorescent immunohistochemistry with identical species-derived primary antibodies. J. Histochem. Cytochem. 2004 52 1219-1230. 

Figure 7.7 Peptide spot color intensity as a function of doubling dilutions of primary (PR) antibody. PR peptides and PR+ tissue sections were both placed on the same slides and stained with various dilutions of the PR mAb. Color intensity of the peptide spots (square symbols) or tumor cells (triangle symbols) was measured and plotted on the y-axis. The figure shows a linear decline in intensity with decreasing antibody concentrations for both the peptide spots and the tissue sections. Tissue color intensity is measured as optical density on a 0-2 scale. Peptide spot color is measured as mean pixel intensity on a 1-256 scale. Copied with permission from Sompuram et al.6...

Double-label ICC is another approach using a similar principle. It pairs a nuclear antibody with a cytoplasmic antibody on the same sample, such as keratin and estrogen receptor. At present, this method is mostly used in the research field. 

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