Antibody Control for the Same Species with Zenon

Fab-labeled reagents based on the species of the 1° antibody and the fluorophore needed. 

Finally, to block the excess labeled Fab reagent molecules, add 10 til of normal mouse serum containing IgG to the above solution for a second incubation. The amount of normal mouse serum by weight should be five times that of the 1° antibody. When 2 pig of 1° antibody is used then 10 fig of normal mouse serum is needed. Each individual 1° antibody must be incubated with the labeled Fab reagent and the blocking normal mouse serum before being combined into a single antibody incubation solution. 

The most common problems with this method are that little or no labeling is seen with the final labeled antibody rather than too much labeling. Correct this situation by adding more labeled Fab reagent to the E antibody and increase the ratio from 6 1 to 10 1. Changing the ratio will require that the control experiments be performed with antibody labeled with the higher ratio. 

Design the Antibody Control for the Same Species with Zenon 

Two antibody controls are needed for labeled Fab E antibody Zenon immunocytochemistry (Table 12.4). These controls will show that each 1° antibody is labeled correctly. To generate a control and additional E antibody labeling reaction is needed, where the blocking normal mouse serum is added to the labeled Fab reagent (Zenon reagent) before the E antibody. In this incubation, the labeled 

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