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文献和实验, incubate for 30 min at 37°C. ( 5g powdered milk in 95 ml PBS)5 Rinse with PBS 3x.6 Add 50 µl of antibody, incubate 30 min at 37°C.7 Rinse with PBS 3x.8 Add 50 µl of goat anti-mouse IgG conjugate with alkaline phosphatase.(1 µl of conjugate in 4 ml of PBS
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
Direct Immunofluorescence Labeling
to 1 million per ml. with culture media or PBS. Place 1 ml. of the cell suspension into each of the 12x75 tubes. You will need a tube for each antibody plus the negative control. If you''re doing simultaneous labeling, use one tube for each combination
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