Your colleagues submitted great questions during the webinar, and we’ve collected and answered them here in the following Q&A:
For the direct conjugate strategy, what would be the proper negative control?
In double- or triple-labeling experiments, is there any problem with incubating all of the primaries in the same step?
Is it possible to combine labeled primary antibodies with other same-species antibodies which would be secondary-labeled with different dyes in a multiplex protocol?
What kind of blocking reagent is good for fluorescent labeling?
Is there a special blocking agent used to block endogenous biotin?
Is there a special blocking agent used to block endogenous peroxidase when using TSA amplification?
Do you suggest using BSA or casein as a blocking solution?
How many dyes per avidin?
For secondary labeling, can sensitivity be increased by using higher concentrations of secondary antibody?
Does my secondary have to be from a species different from that of my tissue?
if you need to use chicken anti-goat and goat anti-rabbit secondaries, is it possible to prevent them from binding to one another?
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