I just want to know that after in vivo antibody injection, the levels of antibody are higher several hours post-injection compared to several days post-injection.
I'm not sure if antibody injection will affect the protein levels, so I don't want to measure that. I'd just like to show that yes the antibody is circulating in the system, and that its levels decrease over time.
Jane Lindborg
Have you tried ELISA assay?
1. coat an ELISA plate with Ag for the Abs you want to detect in sera.
I'd recommend coating over night in 4C with 100ul/well 3ug/ml Ag in PBS
2. wash with 1% PBST
3. plate diluted sera samples (different time points) + required controls. incubate 1 hour
I'd recommend diluting sera 1:500.
4. wash 5 times
5. plate HRP conjugated anti mouse 1 hour
I'd recommend using 100ul/well 3ug/ml of Ab
6. wash 5 times
7. plate TMB 100ul/well
8. read OD in 650nm
