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文献和实验Immunofluorescent Staining of Mouse and Rat Leukocytes
receptors), a three-step protocol may amplify the staining: use purified primary antibody (steps 2-4 of above procedure), biotinylated anti-Ig for the 2nd step (steps 6-7, above), and fluorochrome-conjugated avidin or streptavidin as the 3rd step (repeat
Cell Surface Immunofluorescence Staining Protocol
of desired fluorochrome conjugated, biotinylated, or purified primary antibodies to 100 μl of anti-coagulated whole blood. 2) Incubate at room temperature for 15-20 minutes in the dark. 3) Dilute 10X Red Blood Cell (RBC) Lysis Buffer
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
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