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ELISA conditions such as coating buffer and blocking solution were optimized using sera obtained from mice immunized with amphotropic MLV particles.
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To optimize the protocol, individual parameters (i.e., filtration, fixation, EDTA addition, staining temperature, and dilution solution) were optimized while keeping the other parameter fixed.
Biotin blocking solution was added, samples were incubated and centrifuged.
Ultrablock (blocking solution) was from Serotec (Raleigh, NC).
The excess blocking solution was removed with filter paper.
Primary antibody (in blocking solution) was incubated for 60 min and then washed three times with blocking solution.
Ionic blocking solutions were used to isolate capacitive currents.
Blocking solution was removed and primary antibody in blocking solution applied at the required dilution.
Blocking solution was used to block the endogenous peroxidase activity.
The parameters of the MS were optimized using a standard solution.
The MS/MS conditions were optimized by manual tuning during pump-infusion of neat solutions.
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