Tue Feb 15, 2022 7:46 pm by mleipold
Hi Natalia,
You didn't say whether you've been successful with this particular antibody previously.
Unfortunately, sometimes a given conjugation just doesn't work.
1. Bad lot of antibody from the manufacturer. It's rare, but I've had a bad lot from even big companies like BD and Biolegend: no problem with a product for years, then 2-3 failed conjugations from the same lot, and then no problem once I switched to another lot.
2. Hole in the filter and your antibody is lost in the flow-through. Again, rare, but does happen.
3. Antibody can't survive the reduction process and precipitates. This should be a repeated error (ie, it never works).
4. Didn't fully redissolve if supplied as lyophilized powder. The CCR7 clone I use comes as a powder, and I had bad recovery on it for a long time until I spent extra time letting the powder dissolve (several pipet to mix and wait, pipet to mix and wait steps).
Mike