First the objective was to
1) Capture L1 RNP from DC and PC labelled cells from Spain, however we had some issues detecting exogenous/endogenous ORF1/ORF2 in these cells (protein expression seemed to be much lower than in MT302 or LD401 HEK cells).
Finally we could detect ORF1 (endogenous+exogenous) so we decided to try to capture endogenous ORF1, but this material was thought for another purpose and using it wouldn't have been clean - powder is stored in case we could use it for something else in the future.
From this point, we have different proyect lines:
1) Capture L1 RNP from DC and PC labelled cells from Spain -
(Given up)
2) Capture endogenous L1 from PA-1 cells - Ongoing
3) Tet-ON PA-1 cells
To get cell lines where we could induce the expression of L1 constructs (and get higher expression levels)
5) Wes LINE-1 antibodies/IPs optimization + Wes quantitative analysis + Combined ORF1/ORF2 Wes
To be able to use Wes for L1 samples, and to know the relative amount of protein in our samples.