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文献和实验免疫组化技术-Immunohistochemical Protocols
Incubation in affinity purified indocarbocyanine (Cy3, viewed in red field, 1:1000 in BSA diluent) or fluorescein isothiocyanate (FITC, viewed in green field, 1:100 in BSA diluent) fluorochrome conjugated secondary antibodies (F[ab']2 fragments
Dual- and Triple-Color Fluorospot
for the other cytokine (Fig. 1). As a second step in the detection, streptavidin conjugated to a red fluorophore (Cy3) and anti-FITC antibodies labeled with a green fluorophore, respectively, are used (10, 11). The resulting spots are subsequently analyzed using
at 1:200 dilution) Cy3-conjugated affinity-purified goat anti-mouse IgG (Jackson Labs; 1 mg/ml stock used at 1:300 dilution) Texas Red-conjugated affinity-purified donkey anti-rabbit (Jackson Labs;1 mg/ml stock used at 1:200 dilution)
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