I ordered the Streptavidin-ZAP (Cat. #IT-27) and had my antibody biotinylated a couple of months ago. I am ready to begin the first round of experiments to determine the concentration needed for the secondary. How much of the biotinylated antibody should I put to combine with the streptavidin for intravitreal injections? Can you please send me a protocol for how to determine the ratio of primary to secondary?
The streptavidin-ZAP should be mixed with the biotinylated material at an equimolar concentration. The streptavidin-ZAP you ordered should have included a data sheet which gives the protein concentration and molecular weight, which you would use to determine the molar concentration. We have a calculator page on our website which can help with this if needed.
Here is a link to our references page for Streptavidin-ZAP which lists the different publications using this product, and I have also listed a specific publication using streptavidin-ZAP combined with a biotinylated antibody being used in intravitreal injections. The reference describes in detail the quantities they tried.
Ren C, Luan L, Wui-Man Lau B, Huang X, Yang J, Zhou Y, Wu X, Gao J, Pickard GE, So KF, Pu M. (2013) Direct Retino-Raphe Projection Alters Serotonergic Tone and Affective Behavior. Neuropsychopharmacology 38(7):1163-1175.
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