with lectins conjugated with TRITC?
Answer. The general rule of thumb when staining with fluorescent protein conjugates is to bracket around 10 micrograms per mL. When using a good fluorescent IgG conjugate, I found that 5 micrograms/mL was a bit dim, whereas 20 micrograms/mL often had a bit too much background. This rule of thumb depends somewhat on the fluorophore (some yield a higher background, etc), but for TRITC conjugates, 10 micrograms/mL usually works well. Although the molecular weight of your lectin is probably is a bit less than IgG, a 2-3 fold difference in molecular weight prabably won’t make that much of a difference. I used to use a TRITC conjugate of wheat germ agglutinin at 10 micrograms per mL and it stained beautifully. Karen Larison, in Oregon (larisonk[AT]uoneuro.uoregon.
Answer. The general rule of thumb when staining with fluorescent protein conjugates is to bracket around 10 micrograms per mL. When using a good fluorescent IgG conjugate, I found that 5 micrograms/mL was a bit dim, whereas 20 micrograms/mL often had a bit too much background. This rule of thumb depends somewhat on the fluorophore (some yield a higher background, etc), but for TRITC conjugates, 10 micrograms/mL usually works well. Although the molecular weight of your lectin is probably is a bit less than IgG, a 2-3 fold difference in molecular weight prabably won’t make that much of a difference. I used to use a TRITC conjugate of wheat germ agglutinin at 10 micrograms per mL and it stained beautifully.
