Dynabeads Protein

Figure 8.1 The results of IHC of two experiments using Dynabeads (Dynal, New York, NY) coated with biotinylated anti-mouse IgG (first experiment) and protein S-100 (second experiment), (a) Positive control showing red color (S-100) localized in the melanoma cells, (b) Strong positive red color circles all beads coated with biotinylated anti-mouse antibody after the heating AR treatment (first experiment), (c) Using the heating AR treatment, S-100-coated polymer beads show positive red color around the beads as circles (second experiment), (d) Negative control of the first experiment. No red color could be seen for polymer beads (arrows) that had been treated with exactly the same protocol as that of slide (b), but omitting the avidin-biotin-peroxidase (label). Bar = 50pm. Reproduced with permission from Shi et al., J. Histochem. Cytochem. 2005 53 1167-1170. See color insert.

Figure 8.1 The results of IHC of two experiments using Dynabeads (Dynal, New York, NY) coated with biotinylated anti-mouse IgG (first experiment) and protein S-100 (second experiment). (See text for full caption). 

Tosylactivated magnetic beads (MB-Tosyl) (Dynabeads M-280 Prod. No. 142.03, Dynal Biotech ASA, Oslo, Norway). Hydroquinone (Sigma), hydrogen peroxide (Merck, Germany). Bradford solution (Bio-Rad protein assay Catalog No. 500-0006) was purchased from Bio-Rad laboratories GMGH (Munich, Germany). 

Rat liver membranes presumed to be from Golgi and lysosomal structure were isolated by immunoadsorption on Dynabeads M-500 (Dynal) conjugated to, respectively, anti-API and anti-LAMPl antibodies. After separation of membrane proteins by SDS-PAGE, endogenous ARDl was detected by immunoblotting with specific polyclonal anti-ARDl antibodies (Vitale et al, 1998a). 

Coupling. Wash Dynabeads M-280 Streptavidin three times with buffer A. Add biotinylated proteins in PBS and incubate the mixture at room temperature ft r 30 min, using a ratio of biotinylated proteimbeads of 10 Ag l mg. 

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